U.S. patent application number 11/521887 was filed with the patent office on 2007-03-29 for tail variants of redox-active therapeutics for treatment of mitochondrial diseases and other conditions and modulation of energy biomarkers.
Invention is credited to Sidney M. Hecht, Guy M. Miller.
Application Number | 20070072943 11/521887 |
Document ID | / |
Family ID | 37637229 |
Filed Date | 2007-03-29 |
United States Patent
Application |
20070072943 |
Kind Code |
A1 |
Miller; Guy M. ; et
al. |
March 29, 2007 |
Tail variants of redox-active therapeutics for treatment of
mitochondrial diseases and other conditions and modulation of
energy biomarkers
Abstract
Methods of treating or suppressing mitochondrial diseases, such
as Friedreich's ataxia (FRDA), Leber's Hereditary Optic Neuropathy
(LHON), mitochondrial myopathy, encephalopathy, lactacidosis,
stroke (MELAS), or Kearns-Sayre Syndrome (KSS) are disclosed, as
well as compounds useful in the methods of the invention. Energy
biomarkers useful in assessing the metabolic state of a subject and
the efficacy of treatment are also disclosed.
Inventors: |
Miller; Guy M.; (Carmel,
CA) ; Hecht; Sidney M.; (Charlottesville,
VA) |
Correspondence
Address: |
MORRISON & FOERSTER LLP
755 PAGE MILL RD
PALO ALTO
CA
94304-1018
US
|
Family ID: |
37637229 |
Appl. No.: |
11/521887 |
Filed: |
September 15, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60717678 |
Sep 15, 2005 |
|
|
|
Current U.S.
Class: |
514/519 ;
514/520; 514/690; 514/718; 552/307 |
Current CPC
Class: |
A61P 27/02 20180101;
A61P 25/20 20180101; A61P 25/00 20180101; C07C 50/06 20130101; A61P
25/22 20180101; A61P 25/18 20180101; C07C 50/02 20130101; A61P 3/10
20180101; A61P 27/12 20180101; A61P 3/12 20180101; A61P 43/00
20180101; A61P 25/08 20180101; A61K 31/055 20130101; C07C 46/00
20130101; A61P 25/24 20180101; A61P 25/14 20180101; A61P 21/00
20180101; A61K 31/275 20130101; A61P 3/00 20180101; A61P 15/12
20180101; A61P 35/00 20180101; C07C 46/06 20130101; C07C 46/02
20130101; A61K 31/122 20130101; A61P 11/00 20180101; A61K 31/05
20130101; A61P 25/02 20180101; A61P 25/16 20180101; A61P 27/16
20180101; C07C 37/07 20130101; A61P 31/06 20180101; A61P 9/00
20180101; A61P 21/02 20180101; C07C 39/08 20130101; A61K 31/12
20130101; A61P 13/02 20180101; A61P 25/28 20180101; C07C 41/16
20130101; C07C 41/30 20130101; C07C 46/00 20130101; C07C 50/06
20130101; C07C 46/00 20130101; C07C 50/02 20130101; C07C 46/02
20130101; C07C 50/02 20130101; C07C 46/06 20130101; C07C 50/02
20130101; C07C 41/16 20130101; C07C 43/2055 20130101; C07C 41/30
20130101; C07C 43/225 20130101; C07C 41/30 20130101; C07C 43/2055
20130101; C07C 37/07 20130101; C07C 39/08 20130101 |
Class at
Publication: |
514/519 ;
514/520; 514/690; 514/718; 552/307 |
International
Class: |
A61K 31/275 20060101
A61K031/275; A61K 31/12 20060101 A61K031/12; C07C 50/28 20060101
C07C050/28 |
Claims
1. A method of treating a mitochondrial disorder, modulating one or
more energy biomarkers, normalizing one or more energy biomarkers,
or enhancing one or more energy biomarkers, comprising
administering to a subject a therapeutically effective amount or
effective amount of one or more compounds of the formulas:
##STR20## where R.sub.1, R.sub.2, and R.sub.3 are independently
selected from --C.sub.1-C.sub.4 alkyl, --C.sub.1-C.sub.4 haloalkyl,
--CN, --F, --Cl, --Br, and --I; R.sub.20 is independently selected
from --C.sub.1-C.sub.20 alkyl, --C.sub.1-C.sub.20 alkenyl,
--C.sub.1-C.sub.20 alkynyl, and --C.sub.1-C.sub.20 containing at
least one double bond and at least one triple bond; and all salts,
stereoisomers, mixtures of stereoisomers, prodrugs, metabolites,
solvates, and hydrates thereof.
2. The method of claim 1, with the proviso that R.sub.20 excludes
C.sub.6 n-alkyl, C.sub.7 n-alkyl, and C.sub.11 n-alkyl.
3. The method of claim 1, where R.sub.1 is independently selected
from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl,
isobutyl, sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.1 to
the remainder of the molecule can be at any location on the alkyl
fragment; where R.sub.2 is independently selected from methyl,
ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.2 to
the remainder of the molecule can be at any location on the alkyl
fragment; where R.sub.3 is independently selected from methyl,
ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.3 to
the remainder of the molecule can be at any location on the alkyl
fragment; where R.sub.20 is independently selected from
--C.sub.1-C.sub.20 alkyl, --C.sub.1-C.sub.20 alkenyl,
--C.sub.1-C.sub.20 alkynyl, and --C.sub.1-C.sub.20 containing at
least one double bond and at least one triple bond and all salts,
stereoisomers, mixtures of stereoisomers, prodrugs, metabolites,
solvates, and hydrates thereof.
4. The method of claim 3, with the proviso that R.sub.20 excludes
C.sub.6 n-alkyl, C.sub.7 n-alkyl, and C.sub.11 n-alkyl.
5. The method of claim 1, wherein at least one of R.sub.1, R.sub.2,
and R.sub.3 is not methyl.
6. The method of claim 2, wherein at least one of R.sub.1, R.sub.2,
and R.sub.3 is not methyl.
7. The method of claim 3, wherein at least one of R.sub.1, R.sub.2,
and R.sub.3 is not methyl.
8. The method of claim 4, wherein at least one of R.sub.1, R.sub.2,
and R.sub.3 is not methyl.
9. The method of claim 1, wherein the mitochondrial disorder is
selected from the group consisting of inherited mitochondrial
diseases; Myoclonic Epilepsy with Ragged Red Fibers (MERRF);
Mitochondrial Myopathy, Encephalopathy, Lactacidosis, Stroke
(MELAS); Leber's Hereditary Optic Neuropathy (LHON); Leigh Disease;
Kearns-Sayre Syndrome (KSS); Friedreich's Ataxia (FA); other
myopathies; cardiomyopathy; encephalomyopathy; renal tubular
acidosis; neurodegenerative diseases; Parkinson's disease;
Alzheimer's disease; amyotrophic lateral sclerosis (ALS); motor
neuron diseases; other neurological diseases; epilepsy; genetic
diseases; Huntington's Disease; mood disorders; schizophrenia;
bipolar disorder; age-associated diseases; macular degeneration;
diabetes; and cancer.
10. The method of claim 1, wherein the mitochondrial disorder is
selected from the group consisting of inherited mitochondrial
diseases; Myoclonic Epilepsy with Ragged Red Fibers (MERRF);
Mitochondrial Myopathy, Encephalopathy, Lactacidosis, Stroke
(MELAS); Leber's Hereditary Optic Neuropathy (LHON); Leigh Disease;
Keams-Sayre Syndrome (KSS); and Friedreich's Ataxia (FA).
11. The method of claim 1, wherein the energy biomarker is selected
from the group consisting of: lactic acid (lactate) levels, either
in whole blood, plasma, cerebrospinal fluid, or cerebral
ventricular fluid; pyruvic acid (pyruvate) levels, either in whole
blood, plasma, cerebrospinal fluid, or cerebral ventricular fluid;
lactate/pyruvate ratios, either in whole blood, plasma,
cerebrospinal fluid, or cerebral ventricular fluid; phosphocreatine
levels, NADH (NADH+H.sup.+) levels; NADPH (NADPH+H.sup.+) levels;
NAD levels; NADP levels; ATP levels; reduced coenzyme Q
(CoQ.sup.red) levels; oxidized coenzyme Q (CoQ.sup.ox) levels;
total coenzyme Q (CoQ.sup.tot) levels; oxidized cytochrome C
levels; reduced cytochrome C levels; oxidized cytochrome C/reduced
cytochrome C ratio; acetoacetate levels, .beta.-hydroxy butyrate
levels, acetoacetate/.beta.-hydroxy butyrate ratio,
8-hydroxy-2'-deoxyguanosine (8-OHdG) levels; levels of reactive
oxygen species; levels of oxygen consumption (VO2); levels of
carbon dioxide output (VCO2); respiratory quotient (VCO2/VO2);
exercise tolerance; and anaerobic threshold.
12. The method of claim 1, wherein the subject is selected from the
group consisting of: a subject with a mitochondrial disease; a
subject undergoing strenuous or prolonged physical activity; a
subject with chronic energy problems; a subject with chronic
respiratory problems; a pregnant female; a pregnant female in
labor; a neonate; a premature neonate; a subject exposed to an
extreme environment; a subject exposed to a hot environment; a
subject exposed to a cold environment; a subject exposed to an
environment with lower-than-average oxygen content; a subject
exposed to an environment with higher-than-average carbon dioxide
content; a subject exposed to an environment with
higher-than-average levels of air pollution; a subject with lung
disease; a subject with lower-than-average lung capacity; a
tubercular patient; a lung cancer patient; an emphysema patient; a
cystic fibrosis patient; a subject recovering from surgery; a
subject recovering from illness; a subject undergoing acute trauma;
a subject in shock; a subject requiring acute oxygen
administration; a subject requiring chronic oxygen administration;
an elderly subject; an elderly subject experiencing decreased
energy; and a subject suffering from chronic fatigue.
13. A compound of the formula: ##STR21## where R.sub.1, R.sub.2,
and R.sub.3 are independently selected from --C.sub.1-C.sub.4
alkyl, --C.sub.1-C.sub.4 haloalkyl, --CN, --F, --Cl, --Br, and --I;
R.sub.20 is independently selected from --C.sub.1-C.sub.20 alkyl,
--C.sub.1-C.sub.20 alkenyl, --C.sub.1-C.sub.20 alkynyl, and
--C.sub.1-C.sub.20 containing at least one double bond and at least
one triple bond; with the proviso that R.sub.20 excludes C.sub.6
n-alkyl, C.sub.7 n-alkyl, and C.sub.11 n-alkyl when R.sub.1,
R.sub.2, and R.sub.3 are all methyl and the additional proviso that
R.sub.20 excludes C.sub.6 n-alkyl when R.sub.3 is bromo, one of
R.sub.1 and R.sub.2 is methyl, and the other one of R.sub.1 and
R.sub.2 is bromo; and all salts, stereoisomers, mixtures of
stereoisomers, prodrugs, metabolites, solvates, and hydrates
thereof.
14. A compound of claim 13, where R.sub.20 excludes C.sub.6
n-alkyl, C.sub.7 n-alkyl, and C.sub.11 n-alkyl for any selections
of R.sub.1, R.sub.2, and R.sub.3.
15. A compound of claim 13, where R.sub.1 is independently selected
from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl,
isobutyl, sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.1 to
the remainder of the molecule can be at any location on the alkyl
fragment; where R.sub.2 is independently selected from methyl,
ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.2 to
the remainder of the molecule can be at any location on the alkyl
fragment; and where R.sub.3 is independently selected from methyl,
ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.3 to
the remainder of the molecule can be at any location on the alkyl
fragment; and all salts, stereoisomers, mixtures of stereoisomers,
prodrugs, metabolites, solvates, and hydrates thereof.
16. A compound of claim 13, wherein at least one of R.sub.1,
R.sub.2, and R.sub.3 is not methyl, and all salts, stereoisomers,
mixtures of stereoisomers, prodrugs, metabolites, solvates, and
hydrates thereof.
17. A compound of claim 16, wherein R.sub.1, R.sub.2, and R.sub.3
are independently selected from C.sub.2-C.sub.4 alkyl, and all
salts, stereoisomers, mixtures of stereoisomers, prodrugs,
metabolites, solvates, and hydrates thereof.
18. A compound of claim 13, wherein one and only one of R.sub.1,
R.sub.2, and R.sub.3 is methyl, and all salts, stereoisomers,
mixtures of stereoisomers, prodrugs, metabolites, solvates, and
hydrates thereof.
19. A compound of claim 13, wherein two and only two of R.sub.1,
R.sub.2, and R.sub.3 are methyl, and all salts, stereoisomers,
mixtures of stereoisomers, prodrugs, metabolites, solvates, and
hydrates thereof.
20. A compound of claim 13, wherein all of R.sub.1, R.sub.2, and
R.sub.3 are methyl, and all salts, stereoisomers, mixtures of
stereoisomers, prodrugs, metabolites, solvates, and hydrates
thereof.
21. A compound of claim 13, additionally comprising a
pharmaceutically acceptable carrier.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the priority benefit of U.S.
Provisional Patent Application No. 60/717,678, filed Sep. 15, 2005.
The entire content of that application is hereby incorporated
herein by reference in its entirety.
TECHNICAL FIELD
[0002] The application discloses compositions and methods useful
for treatment or suppression of diseases due to mitochondrial
disorders, such as Friedreich's ataxia, Leber's Hereditary Optic
Neuropathy, Kearns-Sayre Syndrome, and mitochondrial myopathy,
encephalopathy, lactacidosis, stroke (MELAS), and for modulating
energy biomarkers in a subject.
BACKGROUND
[0003] Mitochondria are organelles in eukaryotic cells, popularly
referred to as the "powerhouse" of the cell. The molecule adenosine
triphosphate (ATP) functions as an energy "currency" or energy
carrier in the cell, and eukaryotic cells derive the majority of
their ATP from biochemical processes carried out by mitochondria.
These biochemical processes include the citric acid cycle (the
tricarboxylic acid cycle, or Kreb's cycle), which generates reduced
nicotinamide adenine dinucleotide (NADH+H.sup.+) from oxidized
nicotinamide adenine dinucleotide (NAD.sup.+), and oxidative
phosphorylation, during which NADH+H.sup.+ is oxidized back to
NAD.sup.+. (The citric acid cycle also reduces flavin adenine
dinucleotide, or FAD, to FADH.sub.2; FADH.sub.2 also participates
in oxidative phosphorylation.)
[0004] The electrons released by oxidation of NADH+H.sup.+ are
shuttled down a series of protein complexes (Complex I, Complex II,
Complex III, and Complex IV) known as the respiratory chain. These
complexes are embedded in the inner membrane of the mitochondrion.
Complex IV, at the end of the chain, transfers the electrons to
oxygen, which is reduced to water. The energy released as these
electrons traverse the complexes is used to generate a proton
gradient across the inner membrane of the mitochondrion, which
creates an electrochemical potential across the inner membrane.
Another protein complex, Complex V (which is not directly
associated with Complexes I, II, III and IV) uses the energy stored
by the electrochemical gradient to convert ADP into ATP.
[0005] The citric acid cycle and oxidative phosphorylation are
preceded by glycolysis, in which a molecule of glucose is broken
down into two molecules of pyruvate, with net generation of two
molecules of ATP per molecule of glucose. The pyruvate molecules
then enter the mitochondria, where they are completely oxidized to
CO.sub.2 and H.sub.2O via oxidative phosphorylation (the overall
process is known as aerobic respiration). The complete oxidation of
the two pyruvate molecules to carbon dioxide and water yields about
at least 28-29 molecules of ATP, in addition to the 2 molecules of
ATP generated by transforming glucose into two pyruvate molecules.
If oxygen is not available, the pyruvate molecule does not enter
the mitochondria, but rather is converted to lactate, in the
process of anaerobic respiration.
[0006] The overall net yield per molecule of glucose is thus
approximately at least 30-31 ATP molecules. ATP is used to power,
directly or indirectly, almost every other biochemical reaction in
the cell. Thus, the extra (approximately) at least 28 or 29
molecules of ATP contributed by oxidative phosphorylation during
aerobic respiration are critical to the proper functioning of the
cell. Lack of oxygen prevents aerobic respiration and will result
in eventual death of almost all aerobic organisms; a few organisms,
such as yeast, are able to survive using either aerobic or
anaerobic respiration.
[0007] When cells in an organism are temporarily deprived of
oxygen, anaerobic respiration is utilized until oxygen again
becomes available or the cell dies. The pyruvate generated during
glycolysis is converted to lactate during anaerobic respiration.
The buildup of lactic acid is believed to be responsible for muscle
fatigue during intense periods of activity, when oxygen cannot be
supplied to the muscle cells. When oxygen again becomes available,
the lactate is converted back into pyruvate for use in oxidative
phosphorylation.
[0008] Genetic defects in the proteins making up the respiratory
chain lead to severe disease states. One such disease is
Friedreich's ataxia (FRDA or FA). Friedreich's ataxia is an
autosomal recessive neurodegenerative and cardiodegenerative
disorder caused by decreased levels of the protein frataxin.
Frataxin is important for the assembly of iron-sulfur clusters in
mitochondrial respiratory-chain complexes. Estimates of the
prevalence of FRDA in the United States range from 1 in every
22,000 people (see World-Wide-Web address
.nlm.nih.gov/medlineplus/ency/article/001411.htm) to I in 50,000
people (World-Wide-Web address
.umc-cares.org/health_info/ADAM/Articles/001411.asp). The disease
causes the progressive loss of voluntary motor coordination
(ataxia) and cardiac complications. Symptoms typically begin in
childhood, and the disease progressively worsens as the patient
grows older; patients eventually become wheelchair-bound due to
motor disabilities.
[0009] Another disease linked to mitochondrial dysfunction is
Leber's Hereditary Optic Neuropathy (LHON). The disease is
characterized by blindness which occurs on average between 27 and
34 years of age (World-Wide-Web address
.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=535000); blindness can
develop in both eyes simultaneously, or sequentially (one eye will
develop blindness, followed by the other eye two months later on
average). Other symptoms may also occur, such as cardiac
abnormalities and neurological complications.
[0010] Yet another devastating syndrome resulting from
mitochondrial defects is mitochondrial myopathy, encephalopathy,
lactacidosis, and stroke (MELAS). The disease can manifest itself
in infants, children, or young adults. Strokes, accompanied by
vomiting and seizures, are one of the most serious symptoms; it is
postulated that the metabolic impairment of mitochondria in certain
areas of the brain is responsible for cell death and neurological
lesions, rather than the impairment of blood flow as occurs in
ischemic stroke. Other severe complications, including neurological
symptoms, are often present, and elevated levels of lactic acid in
the blood occur.
[0011] Another mitochondrial disease is Kearns-Sayre Syndrome
(KSS). KSS is characterized by a triad of features including: (1)
typical onset in persons younger than age 20 years; (2) chronic,
progressive, external ophthalmoplegia; and (3) pigmentary
degeneration of the retina. In addition, KSS may include cardiac
conduction defects, cerebellar ataxia, and raised cerebrospinal
fluid (CSF) protein levels (e.g., >100 mg/dL). Additional
features associated with KSS may include myopathy, dystonia,
endocrine abnormalities (e.g., diabetes, growth retardation or
short stature, and hypoparathyroidism), bilateral sensorineural
deafness, dementia, cataracts, and proximal renal tubular acidosis.
Thus, KSS may affect many organ systems.
[0012] The four diseases above appear to be caused by defects in
complex I of the respiratory chain. Electron transfer from complex
I to the remainder of the respiratory chain is mediated by the
compound coenzyme Q (also known as ubiquinone). Oxidized coenzyme Q
(CoQ.sup.ox or ubiquinone) is reduced by complex I to reduced
coenzyme Q (CoQ.sup.red or ubiquinol). The reduced coenzyme Q then
transfers its electrons to complex III of the respiratory chain
(skipping over complex II), where it is re-oxidized to CoQ.sup.ox
(ubiquinone). CoQ.sup.ox can then participate in further iterations
of electron transfer.
[0013] Very few treatments are available for patients suffering
from these diseases. Recently, the compound idebenone has been
proposed for treatment of Friedreich's ataxia. While the clinical
effects of idebenone have been relatively modest, the complications
of mitochondrial diseases can be so severe that even marginally
useful therapies are preferable to the untreated course of the
disease. Another compound, MitoQ, has been proposed for treating
mitochondrial disorders (see U.S. Patent Application Publication
No. 2005/0043553); clinical results for MitoQ have not yet been
reported. For KSS, administration of coenzyme Q10 (CoQ10) and
vitamin supplements have shown only transient beneficial effects in
individual cases.
[0014] Accordingly, there is a serious and unmet need for effective
treatments of mitochondrial disorders, such as Friedreich's ataxia,
Leber's hereditary optic neuropathy, MELAS, and Kearns-Sayre
Syndrome.
[0015] The ability to adjust biological production of energy has
applications beyond the diseases described above. Various other
disorders can result in suboptimal levels of energy biomarkers
(sometimes also referred to as indicators of energetic function),
such as ATP levels. Treatments for these disorders are also needed,
in order to modulate one or more energy biomarkers to improve the
health of the patient. In other applications, it can be desirable
to modulate certain energy biomarkers away from their normal values
in an individual that is not suffering from disease. For example,
if an individual is undergoing an extremely strenuous undertaking,
it can be desirable to raise the level of ATP in that
individual.
DISCLOSURE OF THE INVENTION
[0016] In one embodiment, the compounds are selected from the group
of formula I consisting of: ##STR1## where R.sub.1, R.sub.2, and
R.sub.3 are independently selected from --C.sub.1-C.sub.4 alkyl,
--C.sub.1-C.sub.4 haloalkyl, --CN, --F, --Cl, --Br, and --I; and
R.sub.20 is independently selected from --C.sub.1-C.sub.20 alkyl,
--C.sub.1-C.sub.20 alkenyl, --C.sub.1-C.sub.20 alkynyl, and
--C.sub.1-C.sub.20 containing at least one double bond and at least
one triple bond; and all salts, stereoisomers, mixtures of
stereoisomers, prodrugs, metabolites, solvates, and hydrates
thereof. All R.sub.1, R.sub.2, and R.sub.3 groups may be linear,
branched, or cyclic. R.sub.20 groups may be linear or branched.
C.sub.1-C.sub.20 alkenyl contains at least one double bond.
C.sub.1-C.sub.20 alkynyl contains at least one triple bond.
[0017] In one embodiment of the above-recited compounds of formula
I, the proviso is added that R.sub.20 cannot be C.sub.6 n-alkyl,
C.sub.7 n-alkyl, or C.sub.11 n-alkyl. In another embodiment of the
above-recited compounds of formula I, the proviso is added when
R.sub.1, R.sub.2, and R.sub.3 are all methyl, R.sub.20 cannot be
C.sub.6 n-alkyl, C.sub.7 n-alkyl, or C.sub.11 n-alkyl. In another
embodiment of the above-recited compounds of formula I, the proviso
is added that R.sub.20 excludes C.sub.6 n-alkyl when R.sub.3 is
bromo, one of R.sub.1 and R.sub.2 is methyl, and the other one of
R.sub.1 and R.sub.2 is bromo. Any one, any two, or all three of
these provisos can also be added to any embodiment of formula I
described herein.
[0018] In another embodiment, the invention embraces a method of
treating or suppressing a mitochondrial disorder, modulating one or
more energy biomarkers, normalizing one or more energy biomarkers,
or enhancing one or more energy biomarkers, by administering a
therapeutically effective amount or effective amount of one or more
compounds of formula I as described above.
[0019] In another embodiment, the invention embraces compounds of
formula I where R.sub.1, R.sub.2, and R.sub.3 are independently
selected from --C.sub.1-C.sub.4 alkyl, --C.sub.1-C.sub.4 haloalkyl,
--CN, --F, --Cl, --Br, and --I, with the proviso that at least one
of R.sub.1, R.sub.2, and R.sub.3 is not methyl; and all salts,
stereoisomers, mixtures of stereoisomers, prodrugs, metabolites,
solvates, and hydrates thereof.
[0020] In another embodiment, the invention embraces compounds of
formula I, where R.sub.1 is independently selected from methyl,
ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.1 to
the remainder of the molecule can be at any location on the alkyl
fragment; where R.sub.2 is independently selected from methyl,
ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.2 to
the remainder of the molecule can be at any location on the alkyl
fragment; and where R.sub.3 is independently selected from methyl,
ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of R.sub.3 to
the remainder of the molecule can be at any location on the alkyl
fragment; with the proviso that at least one of R.sub.1, R.sub.2,
and R.sub.3 is not methyl; and all salts, stereoisomers, mixtures
of stereoisomers, prodrugs, metabolites, solvates, and hydrates
thereof.
[0021] In another embodiment, the invention embraces compounds of
formula I where R.sub.1, R.sub.2, and R.sub.3 are independently
selected from methyl, ethyl, n-propyl, and n-butyl, with the
proviso that at least one of R.sub.1, R.sub.2, and R.sub.3 is not
methyl; and all salts, stereoisomers, mixtures of stereoisomers,
prodrugs, metabolites, solvates, and hydrates thereof.
[0022] In another embodiment, the invention embraces compounds of
formula I where R.sub.1, R.sub.2, and R.sub.3 are independently
selected from C.sub.2-C.sub.4 alkyl; and all salts, stereoisomers,
mixtures of stereoisomers, prodrugs, metabolites, solvates, and
hydrates thereof.
[0023] In another embodiment, the invention embraces compounds of
formula I where R.sub.1, R.sub.2, and R.sub.3 are independently
selected from C.sub.2-C.sub.4 n-alkyl; and all salts,
stereoisomers, mixtures of stereoisomers, prodrugs, metabolites,
solvates, and hydrates thereof.
[0024] In another embodiment, the invention embraces compounds of
formula I, where R.sub.1 is independently selected from ethyl,
n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl,
t-butyl, cyclobutyl, cyclopropyl-methyl, and methyl-cyclopropane,
where the point of attachment of R.sub.1 to the remainder of the
molecule can be at any location on the alkyl fragment; where
R.sub.2 is independently selected from ethyl, n-propyl, isopropyl,
cyclopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, cyclobutyl,
cyclopropyl-methyl, and methyl-cyclopropane, where the point of
attachment of R.sub.2 to the remainder of the molecule can be at
any location on the alkyl fragment; and where R.sub.3 is
independently selected from ethyl, n-propyl, isopropyl,
cyclopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, cyclobutyl,
cyclopropyl-methyl, and methyl-cyclopropane, where the point of
attachment of R.sub.3 to the remainder of the molecule can be at
any location on the alkyl fragment; and all salts, stereoisomers,
mixtures of stereoisomers, prodrugs, metabolites, solvates, and
hydrates thereof.
[0025] In another embodiment, the invention embraces compounds of
formula I wherein any one of R.sub.1, R.sub.2, and R.sub.3 is
methyl and the remaining groups are independently selected from
C.sub.2-C.sub.4 alkyl; and all salts, stereoisomers, mixtures of
stereoisomers, prodrugs, metabolites, solvates, and hydrates
thereof. The C.sub.2-C.sub.4 alkyl groups are independently
selected from ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl,
isobutyl, sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of the
C.sub.2-C.sub.4 alkyl group to the remainder of the molecule can be
at any location on the alkyl fragments.
[0026] In another embodiment, the invention embraces compounds of
formula I wherein any two of R.sub.1, R.sub.2, and R.sub.3 are
methyl and the remaining group is independently selected from
C.sub.2-C.sub.4 alkyl; and all salts, stereoisomers, mixtures of
stereoisomers, prodrugs, metabolites, solvates, and hydrates
thereof. The C.sub.2-C.sub.4 alkyl group is independently selected
from ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl,
sec-butyl, t-butyl, cyclobutyl, cyclopropyl-methyl, and
methyl-cyclopropane, where the point of attachment of the
C.sub.2-C.sub.4 alkyl group to the remainder of the molecule can be
at any location on the alkyl fragment.
[0027] In another embodiment, the invention embraces compounds of
formula I wherein R.sub.1, R.sub.2, and R.sub.3 are all methyl,
with the proviso that R.sub.20 cannot be C.sub.6 n-alkyl, C.sub.7
n-alkyl, or C.sub.11 n-alkyl. In an additional embodiment with this
proviso, one and only one of R.sub.1, R.sub.2, and R.sub.3 is
methyl. In an additional embodiment with this proviso, two and only
two of R.sub.1, R.sub.2, and R.sub.3 are methyl. In an additional
embodiment with this proviso, all three of R.sub.1, R.sub.2, and
R.sub.3 are methyl.
[0028] In another variation, in any of the embodiments of the
compounds of formula I, alkenyl can include adjacent sites of
unsaturation (e.g., allenyl, of the form --C.dbd.C.dbd.C--). In
another variation, in any of the embodiments of the compounds of
formula I, alkenyl excludes adjacent sites of unsaturation such as
allenyl.
[0029] In other embodiments, including any of the foregoing
embodiments, the mitochondrial disorder is selected from the group
consisting of inherited mitochondrial diseases; Myoclonic Epilepsy
with Ragged Red Fibers (MERRF); Mitochondrial Myopathy,
Encephalopathy, Lactacidosis, Stroke (MELAS); Leber's Hereditary
Optic Neuropathy (LHON); Leigh Disease; Kearns-Sayre Syndrome
(KSS); Friedreich's Ataxia (FA); other myopathies; cardiomyopathy;
encephalomyopathy; renal tubular acidosis; neurodegenerative
diseases; Parkinson's disease; Alzheimer's disease; amyotrophic
lateral sclerosis (ALS); motor neuron diseases; other neurological
diseases; epilepsy; genetic diseases; Huntington's Disease; mood
disorders; schizophrenia; bipolar disorder; age-associated
diseases; macular degeneration; diabetes; and cancer.
[0030] In another embodiment, including any of the foregoing
embodiments, the mitochondrial disorder is selected from the group
consisting of inherited mitochondrial diseases; Myoclonic Epilepsy
with Ragged Red Fibers (MERRF); Mitochondrial Myopathy,
Encephalopathy, Lactacidosis, Stroke (MELAS); Leber's Hereditary
Optic Neuropathy (LHON); Leigh Disease; Kearns-Sayre Syndrome
(KSS); and Friedreich's Ataxia (FA).
[0031] In another embodiment of the invention, including any of the
foregoing embodiments, the mitochondrial disorder is Friedreich's
ataxia (FRDA). In another embodiment of the invention, the
mitochondrial disorder is Leber's Hereditary Optic Neuropathy
(LHON). In another embodiment of the invention, the mitochondrial
disorder is mitochondrial myopathy, encephalopathy, lactacidosis,
stroke (MELAS). In another embodiment of the invention, the
mitochondrial disorder is Kearns-Sayre Syndrome (KSS). In another
embodiment of the invention, the mitochondrial disorder is
Myoclonic Epilepsy with Ragged Red Fibers (MERRF). In another
embodiment of the invention, the mitochondrial disorder is
Parkinson's disease.
[0032] In another embodiment of the invention, including any of the
foregoing embodiments, the compounds described herein are
administered to subjects suffering from a mitochondrial disorder to
modulate one or more of various energy biomarkers, including, but
not limited to, lactic acid (lactate) levels, either in whole
blood, plasma, cerebrospinal fluid, or cerebral ventricular fluid;
pyruvic acid (pyruvate) levels, either in whole blood, plasma,
cerebrospinal fluid, or cerebral ventricular fluid;
lactate/pyruvate ratios, either in whole blood, plasma,
cerebrospinal fluid, or cerebral ventricular fluid; phosphocreatine
levels, NADH (NADH+H.sup.+) or NADPH (NADPH+H.sup.+) levels; NAD or
NADP levels; ATP levels; reduced coenzyme Q (CoQ.sup.red) levels;
oxidized coenzyme Q (CoQ.sup.ox) levels; total coenzyme Q
(CoQ.sup.tot) levels; oxidized cytochrome C levels; reduced
cytochrome C levels; oxidized cytochrome C/reduced cytochrome C
ratio; acetoacetate levels; beta-hydroxy butyrate levels;
acetoacetate/beta-hydroxy butyrate ratio;
8-hydroxy-2'-deoxyguanosine (8-OHdG) levels; levels of reactive
oxygen species; oxygen consumption (VO2), carbon dioxide output
(VCO2), respiratory quotient (VCO2/VO2), and to modulate exercise
intolerance (or conversely, modulate exercise tolerance) and to
modulate anaerobic threshold. Energy biomarkers can be measured in
whole blood, plasma, cerebrospinal fluid, cerebroventricular fluid,
arterial blood, venous blood, or any other body fluid, body gas, or
other biological sample useful for such measurement. In one
embodiment, the levels are modulated to a value within about 2
standard deviations of the value in a healthy subject. In another
embodiment, the levels are modulated to a value within about 1
standard deviation of the value in a healthy subject. In another
embodiment, the levels in a subject are changed by at least about
10% above or below the level in the subject prior to modulation. In
another embodiment, the levels are changed by at least about 20%
above or below the level in the subject prior to modulation. In
another embodiment, the levels are changed by at least about 30%
above or below the level in the subject prior to modulation. In
another embodiment, the levels are changed by at least about 40%
above or below the level in the subject prior to modulation. In
another embodiment, the levels are changed by at least about 50%
above or below the level in the subject prior to modulation. In
another embodiment, the levels are changed by at least about 75%
above or below the level in the subject prior to modulation. In
another embodiment, the levels are changed by at least about 100%
above or at least about 90% below the level in the subject prior to
modulation.
[0033] In another embodiment, including any of the foregoing
embodiments, the subject or subjects in which a method of treating
or suppressing a mitochondrial disorder, modulating one or more
energy biomarkers, normalizing one or more energy biomarkers, or
enhancing one or more energy biomarkers is performed is/are
selected from the group consisting of subjects undergoing strenuous
or prolonged physical activity; subjects with chronic energy
problems; subjects with chronic respiratory problems; pregnant
females; pregnant females in labor; neonates; premature neonates;
subjects exposed to extreme environments; subjects exposed to hot
environments; subjects exposed to cold environments; subjects
exposed to environments with lower-than-average oxygen content;
subjects exposed to environments with higher-than-average carbon
dioxide content; subjects exposed to environments with
higher-than-average levels of air pollution; airline travelers;
flight attendants; subjects at elevated altitudes; subjects living
in cities with lower-than-average air quality; subjects working in
enclosed environments where air quality is degraded; subjects with
lung diseases; subjects with lower-than-average lung capacity;
tubercular patients; lung cancer patients; emphysema patients;
cystic fibrosis patients; subjects recovering from surgery;
subjects recovering from illness; elderly subjects; elderly
subjects experiencing decreased energy; subjects suffering from
chronic fatigue; subjects suffering from chronic fatigue syndrome;
subjects undergoing acute trauma; subjects in shock; subjects
requiring acute oxygen administration; subjects requiring chronic
oxygen administration; or other subjects with acute, chronic, or
ongoing energy demands who can benefit from enhancement of energy
biomarkers.
[0034] In another embodiment, the invention embraces one or more
compounds of formula I in combination with a pharmaceutically
acceptable excipient, carrier, or vehicle.
[0035] In another embodiment, the invention embraces the use of one
or more compounds of formula I in therapy. In another embodiment,
the invention embraces the use of one or more compounds of formula
I in the therapy of mitochondrial disease. In another embodiment,
the invention embraces the use of one or more compounds of formula
I in the manufacture of a medicament for use in therapy of
mitochondrial disease.
[0036] For all of the compounds and methods described above, the
quinone form can also be used in its reduced (hydroquinone) form
when desired. Likewise, the hydroquinone form can also be used in
its oxidized (quinone) form when desired. The phrase "compounds of
formula (I)" is intended to include both the oxidized and reduced
form of the compounds, unless otherwise specified.
MODES FOR CARRYING OUT THE INVENTION
[0037] The invention embraces compounds useful in treating or
suppressing mitochondrial disorders, and methods of using such
compounds for modulation of energy biomarkers. The redox active
therapeutics for treatment or suppression of mitochondrial diseases
and associated aspects of the invention are described in more
detail herein.
[0038] By "subject," "individual," or "patient" is meant an
individual organism, preferably a vertebrate, more preferably a
mammal, most preferably a human.
[0039] "Treating" a disease with the compounds and methods
discussed herein is defined as administering one or more of the
compounds discussed herein, with or without additional therapeutic
agents, in order to reduce or eliminate either the disease or one
or more symptoms of the disease, or to retard the progression of
the disease or of one or more symptoms of the disease, or to reduce
the severity of the disease or of one or more symptoms of the
disease. "Suppression" of a disease with the compounds and methods
discussed herein is defined as administering one or more of the
compounds discussed herein, with or without additional therapeutic
agents, in order to suppress the clinical manifestation of the
disease, or to suppress the manifestation of adverse symptoms of
the disease. The distinction between treatment and suppression is
that treatment occurs after adverse symptoms of the disease are
manifest in a subject, while suppression occurs before adverse
symptoms of the disease are manifest in a subject. Suppression may
be partial, substantially total, or total. Because many of the
mitochondrial disorders are inherited, genetic screening can be
used to identify patients at risk of the disease. The compounds
disclosed herein and methods of the invention can then be
administered to or practiced on asymptomatic patients at risk of
developing the clinical symptoms of the disease, in order to
suppress the appearance of any adverse symptoms. "Therapeutic use"
of the compounds discussed herein is defined as using one or more
of the compounds discussed herein to treat or suppress a disease,
as defined above. An "effective amount" of a compound is an amount
of the compound sufficient to modulate, normalize, or enhance one
or more energy biomarkers (where modulation, normalization, and
enhancement are defined below). A "therapeutically effective
amount" of a compound is an amount of the compound, which, when
administered to a subject, is sufficient to reduce or eliminate
either a disease or one or more symptoms of a disease, or to retard
the progression of a disease or of one or more symptoms of a
disease, or to reduce the severity of a disease or of one or more
symptoms of a disease, or to suppress the clinical manifestation of
a disease, or to suppress the manifestation of adverse symptoms of
a disease. A therapeutically effective amount can be given in one
or more administrations. An "effective amount" of a compound
embraces both a therapeutically effective amount, as well as an
amount effective to modulate, normalize, or enhance one or more
energy biomarkers in a subject.
[0040] "Modulation" of, or to "modulate," an energy biomarker means
to change the level of the energy biomarker towards a desired
value, or to change the level of the energy biomarker in a desired
direction (e.g., increase or decrease). Modulation can include, but
is not limited to, normalization and enhancement as defined
below.
[0041] "Normalization" of, or to "normalize," an energy biomarker
is defined as changing the level of the energy biomarker from a
pathological value towards a normal value, where the normal value
of the energy biomarker can be 1) the level of the energy biomarker
in a healthy person or subject, or 2) a level of the energy
biomarker that alleviates one or more undesirable symptoms in the
person or subject. That is, to normalize an energy biomarker which
is depressed in a disease state means to increase the level of the
energy biomarker towards the normal (healthy) value or towards a
value which alleviates an undesirable symptom; to normalize an
energy biomarker which is elevated in a disease state means to
decrease the level of the energy biomarker towards the normal
(healthy) value or towards a value which alleviates an undesirable
symptom.
[0042] "Enhancement" of, or to "enhance," energy biomarkers means
to intentionally change the level of one or more energy biomarkers
away from either the normal value, or the value before enhancement,
in order to achieve a beneficial or desired effect. For example, in
a situation where significant energy demands are placed on a
subject, it may be desirable to increase the level of ATP in that
subject to a level above the normal level of ATP in that subject.
Enhancement can also be of beneficial effect in a subject suffering
from a disease or pathology such as a mitochondrial disease, in
that normalizing an energy biomarker may not achieve the optimum
outcome for the subject; in such cases, enhancement of one or more
energy biomarkers can be beneficial, for example,
higher-than-normal levels of ATP, or lower-than-normal levels of
lactic acid (lactate) can be beneficial to such a subject.
[0043] By modulating, normalizing, or enhancing the energy
biomarker Coenzyme Q is meant modulating, normalizing, or enhancing
the variant or variants of Coenzyme Q which is predominant in the
species of interest. For example, the variant of Coenzyme Q which
predominates in humans is Coenzyme Q10. If a species or subject has
more than one variant of Coenzyme Q present in significant amounts
(i.e., present in amounts which, when modulated, normalized, or
enhanced, can have a beneficial effect on the species or subject),
modulating, normalizing, or enhancing Coenzyme Q can refer to
modulating, normalizing or enhancing any or all variants of
Coenzyme Q present in the species or subject.
[0044] While the compounds described herein can occur and can be
used as the neutral (non-salt) compound, the description is
intended to embrace all salts of the compounds described herein in
addition to the non-salt compounds, as well as methods of using
such salts of the compounds. In one embodiment, the salts of the
compounds comprise pharmaceutically acceptable salts.
Pharmaceutically acceptable salts are those salts which can be
administered as drugs or pharmaceuticals to humans and/or animals
and which, upon administration, retain at least some of the
biological activity of the free compound (neutral compound or
non-salt compound). The desired salt of a basic compound may be
prepared by methods known to those of skill in the art by treating
the compound with an acid. Examples of inorganic acids include, but
are not limited to, hydrochloric acid, hydrobromic acid, sulfinuric
acid, nitric acid, and phosphoric acid. Examples of organic acids
include, but are not limited to, formic acid, acetic acid,
propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic
acid, malonic acid, succinic acid, fumaric acid, tartaric acid,
citric acid, benzoic acid, cinnamic acid, mandelic acid, sulfonic
acids, and salicylic acid. Salts of basic compounds with amino
acids, such as aspartate salts and glutamate salts, can also be
prepared. The desired salt of an acidic compound can be prepared by
methods known to those of skill in the art by treating the compound
with a base. Examples of inorganic salts of acid compounds include,
but are not limited to, alkali metal and alkaline earth salts, such
as sodium salts, potassium salts, magnesium salts, and calcium
salts; ammonium salts; and aluminum salts. Examples of organic
salts of acid compounds include, but are not limited to, procaine,
dibenzylamine, N-ethylpiperidine, N,N'-dibenzylethylenediamine, and
triethylamine salts. Salts of acidic compounds with amino acids,
such as lysine salts, can also be prepared.
[0045] The invention also includes all stereoisomers of the
compounds, including diastereomers and enantiomers. The invention
also includes mixtures of stereoisomers in any ratio, including,
but not limited to, racemic mixtures. Unless stereochemistry is
explicitly indicated in a structure, the structure is intended to
embrace all possible stereoisomers of the compound depicted. If
stereochemistry is explicitly indicated for one portion or portions
of a molecule, but not for another portion or portions of a
molecule, the structure is intended to embrace all possible
stereoisomers for the portion or portions where stereochemistry is
not explicitly indicated.
[0046] The compounds can be administered in prodrug form. Prodrugs
are derivatives of the compounds which are themselves relatively
inactive, but which convert into the active compound when
introduced into the subject in which they are used, by a chemical
or biological process in vivo, such as an enzymatic conversion.
Suitable prodrug formulations include, but are not limited to,
peptide conjugates of the compounds disclosed herein and esters of
compounds disclosed herein. Further discussion of suitable prodrugs
is provided in H. Bundgaard, Design of Prodrugs, New York:
Elsevier, 1985; in R. Silverman, The Organic Chemistry of Drug
Design and Drug Action, Boston: Elsevier, 2004; in R. L. Juliano
(ed.), Biological Approaches to the Controlled Delivery of Drugs
(Annals of the New York Academy of Sciences, v. 507), New York: New
York Academy of Sciences, 1987; and in E. B. Roche (ed.), Design of
Biopharmaceutical Properties Through Prodrugs and Analogs
(Symposium sponsored by Medicinal Chemistry Section, APhA Academy
of Pharmaceutical Sciences, November 1976 national meeting,
Orlando, Fla.), Washington: The Academy, 1977.
[0047] The various compounds disclosed herein can be administered
either as therapeutic agents in and of themselves, or as prodrugs
which will convert to other therapeutically effective or effective
substances in the body.
[0048] Metabolites of the compounds are also embraced by the
invention. However, metabolites of substances which occur naturally
in subjects are excluded from the claimed compounds of the
invention.
[0049] "C.sub.1-C.sub.4 alkyl" is intended to embrace methyl (Me),
ethyl (Et), propyl (Pr), n-propyl (nPr), isopropyl (iPr), butyl
(Bu), n-butyl (nBu), isobutyl (iBu), sec-butyl (sBu), t-butyl
(tBu), cyclopropyl (cyclPr), cyclobutyl (cyclBu),
cyclopropyl-methyl (cyclPr-Me) and methyl-cyclopropane (Me-cyclPr),
where the C.sub.1-C.sub.4 alkyl groups can be attached via any
valence on the C.sub.1-C.sub.4 alkyl groups.
[0050] "Halogen" or "halo" substituents designates fluoro (--F),
chloro (--Cl), bromo (--Br), and iodo (--I).
[0051] "C.sub.1-C.sub.4 haloalkyl" is intended to embrace any
C.sub.1-C.sub.4 alkyl substituent having at least one halogen
substituent; the halogen can be attached via any valence on the
C.sub.1-C.sub.4 alkyl group. One subset of C.sub.1-C.sub.4
haloalkyl is --CF.sub.3, --CCl.sub.3, --CBr.sub.3, and --CI.sub.3.
Another subset of C.sub.1-C.sub.4 haloalkyl is the subset with
exactly one halogen substituent. Another subset of C.sub.1-C.sub.4
haloalkyl is the subset of C.sub.1-C.sub.4 perhaloalkyl; that is,
C.sub.1-C.sub.4 alkyl with all available valences replaced by
halogens. Another subset of C.sub.1-C.sub.4 haloalkyl is the subset
of C.sub.1-C.sub.4 perfluoroalkyl; that is, C.sub.1-C.sub.4 alkyl
with all available valences replaced by fluorines. Another subset
of C.sub.1-C.sub.4 haloalkyl is the subset of C.sub.1-C.sub.4
perchloroalkyl; that is, C.sub.1-C.sub.4 alkyl with all available
valences replaced by chlorines.
Synthesis of Compounds of Formula I
[0052] Synthesis of the compounds disclosed herein is readily
accomplished by one of skill in the art. A synthesis of
benzoquinone-type compounds is disclosed in U.S. Pat. No.
4,393,075. Other methods of interest are found in U.S. Pat. No.
5,229,385 and U.S. Pat. No. 4,310,465.
[0053] A method of synthesizing compounds of formula I is by
adapting the following synthesis for the compound (105): ##STR2##
which is as follows: ##STR3## where the chemistry for conversion of
duroquinone (101) into
3,6-dimethoxy-1,2,4,5-tetramethyl-1,4-cyclohexadiene (102) is
described in Thomas et al., Journal of Organic Chemistry
51(22):4160 (1986); the chemistry for conversion of
3,6-dimethoxy-1,2,4,5-tetramethyl-1,4-cyclohexadiene (102) into the
3,6-dimethoxy-1-methylene
lithium-2,4,5-trimethyl-1,4-cyclohexadiene (103) intermediate is
described in Hubscher et al., Helvetica Chimica Acta 73(4):1068
(1990); and the chemistry for conversion of the
3,6-dimethoxy-1-alkyl-2,4,5-trimethyl-1,4-cyclohexadiene (104) into
the 2-alkyl-3,5,6-trimethyl-1,4-benzoquinone (105) is described in
Shiraishi et al., Journal of Medicinal Chemistry 32(9):2214 (1989).
It should be noted that, while the reaction is illustrated with
methyl as R.sub.1, R.sub.2, and R.sub.3, other R.sub.1, R.sub.2,
and R.sub.3 substituents can be used at the methyl-substituted
locations on the ring.
[0054] This synthesis can be easily modified to produce compounds
with any combination of saturated, unsaturated and/or branched
hydrocarbon chains by using the appropriate bromo compound, that
is, by using a compound of the formula
Br--(CH.sub.2).sub.3--R.sub.20 for the reaction converting 103 to
104, where R.sub.20 is independently selected from
--C.sub.1-C.sub.20 alkyl, --C.sub.1-C.sub.20 alkenyl,
--C.sub.1-C.sub.20 alkynyl, and --C.sub.1-C.sub.20 containing at
least one double bond and at least one triple bond.
[0055] Another method of making compounds of formula I is by
adapting the following synthesis: ##STR4## where the chemistry of
converting 1,4-hydroxy-2,3,5-trimethylbenzene (110) into
2,3,5-trimethyl-1,4-benzoquinone (111) is described in Pelter et
al., J. Chem. Soc., Perkin Trans. 1, (16), 1891 (1993), the
chemistry of converting the benzoquinone compound (111) into the
2-alkyl-3,5,6-trimethyl-1,4-benzoquinone (105) is described in
Fieser et al., Journal of the American Chemical Society 64(9):2060
(1942), and the chemistry of converting the alkanoyl chloride (113)
into the dialkanoyl peroxide (114) is described in Silbert et al.,
Journal of the American Chemical Society 81(10):2364 (1959). The
following compound (115) ##STR5## can be used to prepare compounds
of formula I via this route, by starting with the appropriate
1,4-dihydroxy-2,3,5-substituted-1,4-benzoquinone and using the
appropriate intermediate (115). Again, while the reaction is
illustrated with methyl as R.sub.1, R.sub.2, and R.sub.3, other
R.sub.1, R.sub.2, and R.sub.3 substituents can be used at the
methyl-substituted locations on the ring.
[0056] Another method of making compounds of formula I is by the
following decarboxylative coupling synthetic method: ##STR6## where
the chemistry of converting 1,4-hydroxy-2,3,5-trimethylbenzene
(110) into 2,3,5-trimethyl-1,4-benzoquinone (111) is described in
Pelter et al., J. Chem. Soc., Perkin Trans. 1, (16), 1891 (1993),
and the chemistry of converting the benzoquinone compound (111)
into the 2-alkyl-3,5,6-trimethyl-1,4-benzoquinone (105) is
described in Asin-Cayuela et al., FEBS Letters 571:9 (2004). As
before, while the reaction is illustrated with methyl as R.sub.1,
R.sub.2, and R.sub.3, other R.sub.1, R.sub.2, and R.sub.3
substituents can be used at the methyl-substituted locations on the
ring.
[0057] Yet another method of making compounds of formula I uses
chemistry adapted from Monte, W. T. and Lindbeck, A. C., Organic
Process Research & Development 5:267-269 (2001), as follows.
The R.sub.1, R.sub.2, R.sub.3-substituted benzenediol is protected
with methyl groups, and then a chloromethyl group is substituted
for hydrogen at the valence on the benzene ring occupied by
hydrogen. ##STR7## The chloromethyl compound is then reacted with a
Grignard reagent of the form R.sub.20--(CH.sub.2).sub.3-MgX (where
X is a Grignard-forming precursor, such as a halogen or metal that
can be transmetallated with magnesium, such as lithium) to form the
compound of formula I (reduced) with protected diols. ##STR8## The
product can be oxidized with concomitant removal of the methyl
ethers to give quinone compounds of the formula I; that compound
can be subsequently reduced with an appropriate reagent (such as
sodium dithionite Na.sub.2S.sub.2O.sub.4) to provide dihydroquinone
compounds of formula I. Interconvertibility of Quinone,
Dihydroquinone Forms
[0058] The quinone and dihydroquinone forms of the compounds
disclosed herein are readily interconverted with appropriate
reagents. For example, the quinone form of a compound can be
reduced to the dihydroquinone form with reducing agents such as
sodium dithionite (Na.sub.2S.sub.2O.sub.4). The hydroquinone form
can be oxidized to the quinone form with oxidizing agents such as
ceric ammonium nitrate or ferric chloride. The quinone and
hydroquinone forms are also readily converted electrochemically, as
is well known in the art. See, e.g., Section 33.4 of Streitweiser
& Heathcock, Introduction to Organic Chemistry, New York:
Macmillan, 1976.
[0059] Accordingly, the compounds of formula I can also be prepared
in reduced form, that is, where the "head group" is a
benzene-1,4-diol moiety instead of a 1,4-benzoquinone. These
compounds are of the following formula I-Red: ##STR9## where
R.sub.1, R.sub.2, R.sub.3, and R.sub.20 are as described for
formula I, and all salts, stereoisomers, solvates and hydrates
thereof.
[0060] When the quinone form is drawn and followed by the phrase
"reduced counterpart thereof" or "reduced form" or the like, the
structure and the subsequent phrase are intended to embrace both
the quinone and hydroquinone. Similarly, when the hydroquinone form
is drawn and followed by the phrase "oxidized counterpart thereof"
or "oxidized form thereof" or the like, the structure and the
subsequent phrase are intended to embrace both the hydroquinone and
quinone.
Diseases Amenable to Treatment or Suppression with Compounds
Disclosed Herein, and Methods of the Invention
[0061] A variety of diseases are believed to be caused or
aggravated by mitochondrial disorders and impaired energy
processing, and can be treated or suppressed using the compounds
disclosed herein, and the methods of the invention. Such diseases
include, but are not limited to, inherited mitochondrial diseases,
such as Myoclonic Epilepsy with Ragged Red Fibers (MERRF),
Mitochondrial Myopathy, Encephalopathy, Lactacidosis, Stroke
(MELAS), Leber's Hereditary Optic Neuropathy (LHON, also referred
to as Leber's Disease, Leber's Optic Atrophy (LOA), or Leber's
Optic Neuropathy (LON)), Leigh Disease or Leigh Syndrome,
Kearns-Sayre Syndrome (KSS), Friedreich's Ataxia (FA), other
myopathies (including cardiomyopathy and encephalomyopathy), and
renal tubular acidosis; neurodegenerative diseases, such as
Parkinson's disease, Alzheimer's disease, amyotrophic lateral
sclerosis (ALS, also known as Lou Gehrig's disease), motor neuron
diseases; other neurological diseases such as epilepsy; genetic
diseases such as Huntington's Disease (which is also a neurological
disease); mood disorders such as schizophrenia and bipolar
disorder; and certain age-associated diseases, particularly
diseases for which CoQ10 has been proposed for treatment, such as
macular degeneration, diabetes, and cancer.
Clinical Assessment of Mitochondrial Dysfunction and Efficacy of
Therapy
[0062] Several readily measurable clinical markers are used to
assess the metabolic state of patients with mitochondrial
disorders. These markers can also be used as indicators of the
efficacy of a given therapy, as the level of a marker is moved from
the pathological value to the healthy value. These clinical markers
include, but are not limited to, one or more of the previously
discussed energy biomarkers, such as lactic acid (lactate) levels,
either in whole blood, plasma, cerebrospinal fluid, or cerebral
ventricular fluid; pyruvic acid (pyruvate) levels, either in whole
blood, plasma, cerebrospinal fluid, or cerebral ventricular fluid;
lactate/pyruvate ratios, either in whole blood, plasma,
cerebrospinal fluid, or cerebral ventricular fluid; phosphocreatine
levels, NADH (NADH+H.sup.+) or NADPH (NADPH+H.sup.+) levels; NAD or
NADP levels; ATP levels; anaerobic threshold; reduced coenzyme Q
(CoQ.sup.red) levels; oxidized coenzyme Q (CoQ.sup.ox) levels;
total coenzyme Q (CoQ.sup.tot) levels; oxidized cytochrome C
levels; reduced cytochrome C levels; oxidized cytochrome C/reduced
cytochrome C ratio; acetoacetate levels, P-hydroxy butyrate levels,
acetoacetate/.beta.-hydroxy butyrate ratio,
8-hydroxy-2'-deoxyguanosine (8-OHdG) levels; levels of reactive
oxygen species; and levels of oxygen consumption (VO2), levels of
carbon dioxide output (VCO2), and respiratory quotient (VCO2/VO2).
Several of these clinical markers are measured routinely in
exercise physiology laboratories, and provide convenient
assessments of the metabolic state of a subject. In one embodiment
of the invention, the level of one or more energy biomarkers in a
patient suffering from a mitochondrial disease, such as
Friedreich's ataxia, Leber's hereditary optic neuropathy, MELAS, or
KSS, is improved to within two standard deviations of the average
level in a healthy subject. In another embodiment of the invention,
the level of one or more of these energy biomarkers in a patient
suffering from a mitochondrial disease, such as Friedreich's
ataxia, Leber's hereditary optic neuropathy, MELAS, or KSS is
improved to within one standard deviation of the average level in a
healthy subject. Exercise intolerance can also be used as an
indicator of the efficacy of a given therapy, where an improvement
in exercise tolerance (i.e., a decrease in exercise intolerance)
indicates efficacy of a given therapy.
[0063] Several metabolic biomarkers have already been used to
evaluate efficacy of CoQ10, and these metabolic biomarkers can be
monitored as energy biomarkers for use in the methods of the
current invention. Pyruvate, a product of the anaerobic metabolism
of glucose, is removed by reduction to lactic acid in an anaerobic
setting or by oxidative metabolism, which is dependent on a
functional mitochondrial respiratory chain. Dysfunction of the
respiratory chain may lead to inadequate removal of lactate and
pyruvate from the circulation and elevated lactate/pyruvate ratios
are observed in mitochondrial cytopathies (see Scriver C R, The
metabolic and molecular bases of inherited disease, 7th ed., New
York: McGraw-Hill, Health Professions Division, 1995; and Munnich
et al., J. Inherit. Metab. Dis. 15(4):448-55 (1992)). Blood
lactate/pyruvate ratio (Chariot et al., Arch. Pathol. Lab. Med.
118(7):695-7 (1994)) is, therefore, widely used as a noninvasive
test for detection of mitochondrial cytopathies (see again Scriver
C R, The metabolic and molecular bases of inherited disease, 7th
ed., New York: McGraw-Hill, Health Professions Division, 1995; and
Munnich et al., J. Inherit. Metab. Dis. 15(4):448-55 (1992)) and
toxic mitochondrial myopathies (Chariot et al., Arthritis Rheum.
37(4):583-6 (1994)). Changes in the redox state of liver
mitochondria can be investigated by measuring the arterial ketone
body ratio (acetoacetate/3-hydroxybutyrate: AKBR) (Ueda et al., J.
Cardiol. 29(2):95-102 (1997)). Urinary excretion of
8-hydroxy-2'-deoxyguanosine (8-OHdG) often has been used as a
biomarker to assess the extent of repair of ROS-induced DNA damage
in both clinical and occupational settings (Erhola et al., FEBS
Lett. 409(2):287-91 (1997); Honda et al., Leuk. Res. 24(6):461-8
(2000); Pilger et al., Free Radic. Res. 35(3):273-80 (2001); Kim et
al. Environ Health Perspect 112(6):666-71 (2004)).
[0064] Magnetic resonance spectroscopy (MRS) has been useful in the
diagnoses of mitochondrial cytopathy by demonstrating elevations in
cerebrospinal fluid (CSF) and cortical white matter lactate using
proton MRS (1H-MRS) (Kaufmann et al., Neurology 62(8):1297-302
(2004)). Phosphorous MRS (31P-MRS) has been used to demonstrate low
levels of cortical phosphocreatine (PCr) (Matthews et al., Ann.
Neurol. 29(4):435-8 (1991)), and a delay in PCr recovery kinetics
following exercise in skeletal muscle (Matthews et al., Ann.
Neurol. 29(4):435-8 (1991); Barbiroli et al., J. Neurol.
242(7):472-7 (1995); Fabrizi et al., J. Neurol. Sci. 137(1):20-7
(1996)). A low skeletal muscle PCr has also been confirmed in
patients with mitochondrial cytopathy by direct biochemical
measurements.
[0065] Exercise testing is particularly helpful as an evaluation
and screening tool in mitochondrial myopathies. One of the hallmark
characteristics of mitochondrial myopathies is a reduction in
maximal whole body oxygen consumption (VO2max) (Taivassalo et al.,.
Brain 126(Pt 2):413-23 (2003)). Given that VO2max is determined by
cardiac output (Qc) and peripheral oxygen extraction
(arterial-venous total oxygen content) difference, some
mitochondrial cytopathies affect cardiac function where delivery
can be altered; however, most mitochondrial myopathies show a
characteristic deficit in peripheral oxygen extraction (A-V O2
difference) and an enhanced oxygen delivery (hyperkinetic
circulation) (Taivassalo et al.,. Brain 126(Pt 2):413-23 (2003)).
This can be demonstrated by a lack of exercise induced
deoxygenation of venous blood with direct AV balance measurements
(Taivassalo et al., Ann. Neurol. 51(1):38-44 (2002)) and
non-invasively by near infrared spectroscopy (Lynch et al., Muscle
Nerve 25(5):664-73 (2002); van Beekvelt et al., Ann. Neurol.
46(4):667-70 (1999)).
[0066] Several of these energy biomarkers are discussed in more
detail as follows. It should be emphasized that, while certain
energy biomarkers are discussed and enumerated herein, the
invention is not limited to modulation, normalization or
enhancement of only these enumerated energy biomarkers.
[0067] Lactic acid (lactate) levels: Mitochondrial dysfunction
typically results in abnormal levels of lactic acid, as pyruvate
levels increase and pyruvate is converted to lactate to maintain
capacity for glycolysis. Mitochondrial dysfunction can also result
in abnormal levels of NADH+H.sup.+, NADPH+H.sup.+, NAD, or NADP, as
the reduced nicotinamide adenine dinucleotides are not efficiently
processed by the respiratory chain. Lactate levels can be measured
by taking samples of appropriate bodily fluids such as whole blood,
plasma, or cerebrospinal fluid. Using magnetic resonance, lactate
levels can be measured in virtually any volume of the body desired,
such as the brain.
[0068] Measurement of cerebral lactic acidosis using magnetic
resonance in MELAS patients is described in Kaufinann et al.,
Neurology 62(8):1297 (2004). Values of the levels of lactic acid in
the lateral ventricles of the brain are presented for two mutations
resulting in MELAS, A3243G and A8344G. Whole blood, plasma, and
cerebrospinal fluid lactate levels can be measured by commercially
available equipment such as the YSI 2300 STAT Plus Glucose &
Lactate Analyzer (YSI Life Sciences, Ohio).
[0069] NAD, NADP, NADH and NADPH levels: Measurement of NAD, NADP,
NADH (NADH+H.sup.+) or NADPH (NADPH+H.sup.+) can be measured by a
variety of fluorescent, enzymatic, or electrochemical techniques,
e.g., the electrochemical assay described in US 2005/0067303.
[0070] Oxygen consumption (vO.sub.2 or VO2), carbon dioxide output
(VCO.sub.2 or VCO2), and respiratory quotient (VCO2/VO2): vO.sub.2
is usually measured either while resting (resting vO.sub.2) or at
maximal exercise intensity (vO.sub.2 max). Optimally, both values
will be measured. However, for severely disabled patients,
measurement of vO.sub.2 max may be impractical. Measurement of both
forms of vO.sub.2 is readily accomplished using standard equipment
from a variety of vendors, e.g. Korr Medical Technologies, Inc.
(Salt Lake City, Utah). VCO2 can also be readily measured, and the
ratio of VCO2 to V02 under the same conditions (VCO2/VO2, either
resting or at maximal exercise intensity) provides the respiratory
quotient (RQ).
[0071] Oxidized Cytochrome C, reduced Cytochrome C, and ratio of
oxidized Cytochrome C to reduced Cytochrome C: Cytochrome C
parameters, such as oxidized cytochrome C levels (Cyt C.sub.ox),
reduced cytochrome C levels (Cyt C.sub.red), and the ratio of
oxidized cytochrome C/reduced cytochrome C ratio (Cyt
C.sub.ox)/(Cyt C.sub.red), can be measured by in vivo near infrared
spectroscopy. See, e.g., Rolfe, P., "In vivo near-infrared
spectroscopy," Annu. Rev. Biomed. Eng. 2:715-54 (2000) and
Strangman et al., "Non-invasive neuroimaging using near-infrared
light" Biol. Psychiatry 52:679-93 (2002).
[0072] Exercise tolerance/Exercise intolerance: Exercise
intolerance is defined as "the reduced ability to perform
activities that involve dynamic movement of large skeletal muscles
because of symptoms of dyspnea or fatigue" (Pina et al.,
Circulation 107:1210 (2003)). Exercise intolerance is often
accompanied by myoglobinuria, due to breakdown of muscle tissue and
subsequent excretion of muscle myoglobin in the urine. Various
measures of exercise intolerance can be used, such as time spent
walking or running on a treadmill before exhaustion, time spent on
an exercise bicycle (stationary bicycle) before exhaustion, and the
like. Treatment with the compounds disclosed herein and the methods
of the invention can result in about a 10% or greater improvement
in exercise tolerance (for example, about a 10% or greater increase
in time to exhaustion, e.g. from 10 minutes to 11 minutes), about a
20% or greater improvement in exercise tolerance, about a 30% or
greater improvement in exercise tolerance, about a 40% or greater
improvement in exercise tolerance, about a 50% or greater
improvement in exercise tolerance, about a 75% or greater
improvement in exercise tolerance, or about a 100% or greater
improvement in exercise tolerance. While exercise tolerance is not,
strictly speaking, an energy biomarker, for the purposes of the
invention, modulation, normalization, or enhancement of energy
biomarkers includes modulation, normalization, or enhancement of
exercise tolerance.
[0073] Similarly, tests for normal and abnormal values of pyruvic
acid (pyruvate) levels, lactate/pyruvate ratio, ATP levels,
anaerobic threshold, reduced coenzyme Q (CoQ.sup.red) levels,
oxidized coenzyme Q (CoQ.sup.ox) levels, total coenzyme Q
(CoQ.sup.tot) levels, oxidized cytochrome C levels, reduced
cytochrome C levels, oxidized cytochrome C/reduced cytochrome C
ratio, acetoacetate levels, .beta.-hydroxy butyrate levels,
acetoacetate/.beta.-hydroxy butyrate ratio,
8-hydroxy-2'-deoxyguanosine (8-OHdG) levels, and levels of reactive
oxygen species are known in the art and can be used to evaluate
efficacy of the compounds disclosed herein and the methods of the
invention. (For the purposes of the invention, modulation,
normalization, or enhancement of energy biomarkers includes
modulation, normalization, or enhancement of anaerobic
threshold.)
[0074] Table 1, following, illustrates the effect that various
dysfunctions can have on biochemistry and energy biomarkers. It
also indicates the physical effect (such as a disease symptom or
other effect of the dysfunction) typically associated with a given
dysfunction. It should be noted that any of the energy biomarkers
listed in the table, in addition to energy biomarkers enumerated
elsewhere, can also be modulated, enhanced, or normalized by the
compounds disclosed herein and the methods of the invention.
RQ=respiratory quotient; BMR=basal metabolic rate; HR(CO)=heart
rate (cardiac output); T=body temperature (preferably measured as
core temperature); AT=anaerobic threshold; pH=blood pH (venous
and/or arterial). TABLE-US-00001 TABLE 1 Site of Biochemical
Measurable Physical Dysfunction Event Energy Biomarker Effect
Respiratory .uparw. NADH .DELTA. lactate, Metabolic Chain .DELTA.
lactate: dyscrasia & pyruvate ratio; and fatigue .DELTA.
acetoacetate: .beta.-hydroxy butyrate ratio Respiratory .dwnarw.
H.sup.+ gradient .DELTA. ATP Organ Chain dependent dysfunction
Respiratory .dwnarw. Electron flux .DELTA. VO.sub.2, RQ, BMR,
Metabolic Chain .DELTA.T, AT, pH dyscrasia & fatigue
Mitochondria & .dwnarw. ATP, .dwnarw. VO.sub.2 .DELTA. Work,
.DELTA.HR (CO) Exercise cytosol intolerance Mitochondria &
.dwnarw. ATP .DELTA. PCr Exercise cytosol intolerance Respiratory
.dwnarw. Cyt C.sub.Ox/Red .DELTA. .lamda. .about.700-900 nM
Exercise Chain (Near Infrared intolerance Spectroscopy)
Intermediary .dwnarw. Catabolism .DELTA. C.sup.14-Labeled Metabolic
metabolism substrates dyscrasia & fatigue Respiratory .dwnarw.
Electron flux .DELTA. Mixed Venous VO.sub.2 Metabolic Chain
dyscrasia & fatigue Mitochondria & .uparw. Oxidative
.DELTA. Tocopherol & Uncertain cytosol stress Tocotrienols,
CoQ10, docosahexaenoic acid Mitochondria & .uparw. Oxidative
.DELTA. Glutathione.sub.red Uncertain cytosol stress Mitochondria
& Nucleic acid .DELTA.8-hydroxy Uncertain cytosol oxidation
2-deoxy guanosine Mitochondria & Lipid oxidation
.DELTA.Isoprostane(s), Uncertain cytosol eicosanoids Cell membranes
Lipid oxidation .DELTA.Ethane (breath) Uncertain Cell membranes
Lipid oxidation .DELTA.Malondialdehyde Uncertain
[0075] Treatment of a subject afflicted by a mitochondrial disease
in accordance with the methods of the invention may result in the
inducement of a reduction or alleviation of symptoms in the
subject, e.g., to halt the further progression of the disorder.
[0076] Partial or complete suppression of the mitochondrial disease
can result in a lessening of the severity of one or more of the
symptoms that the subject would otherwise experience. For example,
partial suppression of MELAS could result in reduction in the
number of stroke-like or seizure episodes suffered.
[0077] Any one, or any combination of, the energy biomarkers
described herein provide conveniently measurable benchmarks by
which to gauge the effectiveness of treatment or suppressive
therapy. Additionally, other energy biomarkers are known to those
skilled in the art and can be monitored to evaluate the efficacy of
treatment or suppressive therapy.
Use of Compounds for Modulation of Energy Biomarkers
[0078] In addition to monitoring energy biomarkers to assess the
status of treatment or suppression of mitochondrial diseases, the
compounds disclosed herein can be used in subjects or patients to
modulate one or more energy biomarkers. Modulation of energy
biomarkers can be done to normalize energy biomarkers in a subject,
or to enhance energy biomarkers in a subject.
[0079] Normalization of one or more energy biomarkers is defined as
either restoring the level of one or more such energy biomarkers to
normal or near-normal levels in a subject whose levels of one or
more energy biomarkers show pathological differences from normal
levels (i.e., levels in a healthy subject), or to change the levels
of one or more energy biomarkers to alleviate pathological symptoms
in a subject. Depending on the nature of the energy biomarker, such
levels may show measured values either above or below a normal
value. For example, a pathological lactate level is typically
higher than the lactate level in a normal (i.e., healthy) person,
and a decrease in the level may be desirable. A pathological ATP
level is typically lower than the ATP level in a normal (i.e.,
healthy) person, and an increase in the level of ATP may be
desirable. Accordingly, normalization of energy biomarkers can
involve restoring the level of energy biomarkers to within about at
least two standard deviations of normal in a subject, more
preferably to within about at least one standard deviation of
normal in a subject, to within about at least one-half standard
deviation of normal, or to within about at least one-quarter
standard deviation of normal.
[0080] When an increase in an energy biomarker level is desired to
normalize the one or more such energy biomarker, the level of the
energy biomarker can be increased to within about at least two
standard deviations of normal in a subject, more preferably
increased to within about at least one standard deviation of normal
in a subject, increased to within about at least one-half standard
deviation of normal, or increased to within about at least
one-quarter standard deviation of normal, by administration of one
or more compounds according to the invention. Alternatively, the
level of one or more of the energy biomarkers can be increased by
about at least 10% above the subject's level of the respective one
or more energy biomarkers before administration, by about at least
20% above the subject's level of the respective one or more energy
biomarkers before administration, by about at least 30% above the
subject's level of the respective one or more energy biomarkers
before administration, by about at least 40% above the subject's
level of the respective one or more energy biomarkers before
administration, by about at least 50% above the subject's level of
the respective one or more energy biomarkers before administration,
by about at least 75% above the subject's level of the respective
one or more energy biomarkers before administration, or by about at
least 100% above the subject's level of the respective one or more
energy biomarkers before administration.
[0081] When a decrease in a level of one or more energy biomarkers
is desired to normalize the one or more energy biomarkers, the
level of the one or more energy biomarkers can be decreased to a
level within about at least two standard deviations of normal in a
subject, more preferably decreased to within about at least one
standard deviation of normal in a subject, decreased to within
about at least one-half standard deviation of normal, or decreased
to within about at least one-quarter standard deviation of normal,
by administration of one or more compounds according to the
invention. Alternatively, the level of the one or more energy
biomarkers can be decreased by about at least 10% below the
subject's level of the respective one or more energy biomarkers
before administration, by about at least 20% below the subject's
level of the respective one or more energy biomarkers before
administration, by about at least 30% below the subject's level of
the respective one or more energy biomarkers before administration,
by about at least 40% below the subject's level of the respective
one or more energy biomarkers before administration, by about at
least 50% below the subject's level of the respective one or more
energy biomarkers before administration, by about at least 75%
below the subject's level of the respective one or more energy
biomarkers before administration, or by about at least 90% below
the subject's level of the respective one or more energy biomarkers
before administration.
[0082] Enhancement of the level of one or more energy biomarkers is
defined as changing the extant levels of one or more energy
biomarkers in a subject to a level which provides beneficial or
desired effects for the subject. For example, a person undergoing
strenuous effort or prolonged vigorous physical activity, such as
mountain climbing, could benefit from increased ATP levels or
decreased lactate levels. As described above, normalization of
energy biomarkers may not achieve the optimum state for a subject
with a mitochondrial disease, and such subjects can also benefit
from enhancement of energy biomarkers. Examples of subjects who
could benefit from enhanced levels of one or more energy biomarkers
include, but are not limited to, subjects undergoing strenuous or
prolonged physical activity, subjects with chronic energy problems,
or subjects with chronic respiratory problems. Such subjects
include, but are not limited to, pregnant females, particularly
pregnant females in labor; neonates, particularly premature
neonates; subjects exposed to extreme environments, such as hot
environments (temperatures routinely exceeding about 85-86 degrees
Fahrenheit or about 30 degrees Celsius for about 4 hours daily or
more), cold environments (temperatures routinely below about 32
degrees Fahrenheit or about 0 degrees Celsius for about 4 hours
daily or more), or environments with lower-than-average oxygen
content, higher-than-average carbon dioxide content, or
higher-than-average levels of air pollution (airline travelers,
flight attendants, subjects at elevated altitudes, subjects living
in cities with lower-than-average air quality, subjects working in
enclosed environments where air quality is degraded); subjects with
lung diseases or lower-than-average lung capacity, such as
tubercular patients, lung cancer patients, emphysema patients, and
cystic fibrosis patients; subjects recovering from surgery or
illness; elderly subjects, including elderly subjects experiencing
decreased energy; subjects suffering from chronic fatigue,
including chronic fatigue syndrome; subjects undergoing acute
trauma; subjects in shock; subjects requiring acute oxygen
administration; subjects requiring chronic oxygen administration;
or other subjects with acute, chronic, or ongoing energy demands
who can benefit from enhancement of energy biomarkers.
[0083] Accordingly, when an increase in a level of one or more
energy biomarkers is beneficial to a subject, enhancement of the
one or more energy biomarkers can involve increasing the level of
the respective energy biomarker or energy biomarkers to about at
least one-quarter standard deviation above normal, about at least
one-half standard deviation above normal, about at least one
standard deviation above normal, or about at least two standard
deviations above normal. Alternatively, the level of the one or
more energy biomarkers can be increased by about at least 10% above
the subject's level of the respective one or more energy biomarkers
before enhancement, by about at least 20% above the subject's level
of the respective one or more energy biomarkers before enhancement,
by about at least 30% above the subject's level of the respective
one or more energy biomarkers before enhancement, by about at least
40% above the subject's level of the respective one or more energy
biomarkers before enhancement, by about at least 50% above the
subject's level of the respective one or more energy biomarkers
before enhancement, by about at least 75% above the subject's level
of the respective one or more energy biomarkers before enhancement,
or by about at least 100% above the subject's level of the
respective one or more energy biomarkers before enhancement.
[0084] When a decrease in a level of one or more energy biomarkers
is desired to enhance one or more energy biomarkers, the level of
the one or more energy biomarkers can be decreased by an amount of
about at least one-quarter standard deviation of normal in a
subject, decreased by about at least one-half standard deviation of
normal in a subject, decreased by about at least one standard
deviation of normal in a subject, or decreased by about at least
two standard deviations of normal in a subject. Alternatively, the
level of the one or more energy biomarkers can be decreased by
about at least 10% below the subject's level of the respective one
or more energy biomarkers before enhancement, by about at least 20%
below the subject's level of the respective one or more energy
biomarkers before enhancement, by about at least 30% below the
subject's level of the respective one or more energy biomarkers
before enhancement, by about at least 40% below the subject's level
of the respective one or more energy biomarkers before enhancement,
by about at least 50% below the subject's level of the respective
one or more energy biomarkers before enhancement, by about at least
75% below the subject's level of the respective one or more energy
biomarkers before enhancement, or by about at least 90% below the
subject's level of the respective one or more energy biomarkers
before enhancement.
Use of Compounds in Research Applications, Experimental Systems,
and Assays
[0085] The compounds disclosed herein can also be used in research
applications. For example, a compound disclosed herein can be used
for in vitro, in vivo, or ex vivo experiments to modulate one or
more energy biomarkers in an experimental system. Such experimental
systems can be cell samples, tissue samples, cell components or
mixtures of cell components, partial organs, whole organs, or
organisms. Any one or more of the compounds disclosed herein can be
used in experimental systems or research applications. Such
research applications can include, but are not limited to, use as
assay reagents, elucidation of biochemical pathways, or evaluation
of the effects of other agents on the metabolic state of the
experimental system in the presence/absence of one or more
compounds disclosed herein.
[0086] Additionally, the compounds can be used in biochemical tests
or assays. Such tests can include incubation of one or more
compounds disclosed herein with a tissue or cell sample from a
subject to evaluate a subject's potential response (or the response
of a specific subset of subjects) to administration of said one or
more compounds, or to determine which compound produces the optimum
effect in a specific subject or subset of subjects. One such test
or assay would involve 1) obtaining a cell sample or tissue sample
from a subject in which modulation of one or more energy biomarkers
can be assayed; 2) administering one or more compounds disclosed
herein to the cell sample or tissue sample; and 3) determining the
amount of modulation of the one or more energy biomarkers after
administration of the one or more compounds, compared to the status
of the energy biomarker prior to administration of the one or more
compounds. Another such test or assay would involve 1) obtaining a
cell sample or tissue sample from a subject in which modulation of
one or more energy biomarkers can be assayed; 2) administering at
least two compounds disclosed herein to the cell sample or tissue
sample; 3) determining the amount of modulation of the one or more
energy biomarkers after administration of the at least two
compounds, compared to the status of the energy biomarker prior to
administration of the at least compounds, and 4) selecting a
compound for use in treatment, suppression, or modulation based on
the amount of modulation determined in step 3).
Pharmaceutical Formulations
[0087] The compounds described herein can be formulated as
pharmaceutical compositions by formulation with additives such as
pharmaceutically acceptable excipients, pharmaceutically acceptable
carriers, and pharmaceutically acceptable vehicles. Suitable
pharmaceutically acceptable excipients, carriers and vehicles
include processing agents and drug delivery modifiers and
enhancers, such as, for example, calcium phosphate, magnesium
stearate, talc, monosaccharides, disaccharides, starch, gelatin,
cellulose, methyl cellulose, sodium carboxymethyl cellulose,
dextrose, hydroxypropyl-.beta.-cyclodextrin,
polyvinylpyrrolidinone, low melting waxes, ion exchange resins, and
the like, as well as combinations of any two or more thereof. Other
suitable pharmaceutically acceptable excipients are described in
"Remington's Pharmaceutical Sciences," Mack Pub. Co., New Jersey
(1991), and "Remington: The Science and Practice of Pharmacy,"
Lippincott Williams & Wilkins, Philadelphia, 20th edition
(2003) and 21 st edition (2005), incorporated herein by
reference.
[0088] A pharmaceutical composition can comprise a unit dose
formulation, where the unit dose is a dose sufficient to have a
therapeutic or suppressive effect or an amount effective to
modulate, normalize, or enhance an energy biomarker. The unit dose
may be sufficient as a single dose to have a therapeutic or
suppressive effect or an amount effective to modulate, normalize,
or enhance an energy biomarker. Alternatively, the unit dose may be
a dose administered periodically in a course of treatment or
suppression of a disorder, or to modulate, normalize, or enhance an
energy biomarker.
[0089] Pharmaceutical compositions containing the compounds
disclosed herein may be in any form suitable for the intended
method of administration, including, for example, a solution, a
suspension, or an emulsion. Liquid carriers are typically used in
preparing solutions, suspensions, and emulsions. Liquid carriers
contemplated for use in the practice of the present invention
include, for example, water, saline, pharmaceutically acceptable
organic solvent(s), pharmaceutically acceptable oils or fats, and
the like, as well as mixtures of two or more thereof. The liquid
carrier may contain other suitable pharmaceutically acceptable
additives such as solubilizers, emulsifiers, nutrients, buffers,
preservatives, suspending agents, thickening agents, viscosity
regulators, stabilizers, and the like. Suitable organic solvents
include, for example, monohydric alcohols, such as ethanol, and
polyhydric alcohols, such as glycols. Suitable oils include, for
example, soybean oil, coconut oil, olive oil, safflower oil,
cottonseed oil, and the like. For parenteral administration, the
carrier can also be an oily ester such as ethyl oleate, isopropyl
myristate, and the like. Compositions disclosed herein may also be
in the form of microparticles, microcapsules, liposomal
encapsulates, and the like, as well as combinations of any two or
more thereof.
[0090] Time-release or controlled release delivery systems may be
used, such as a diffusion controlled matrix system or an erodible
system, as described for example in: Lee, "Diffusion-Controlled
Matrix Systems", pp. 155-198 and Ron and Langer, "Erodible
Systems", pp. 199-224, in "Treatise on Controlled Drug Delivery",
A. Kydonieus Ed., Marcel Dekker, Inc., New York 1992. The matrix
may be, for example, a biodegradable material that can degrade
spontaneously in situ and in vivo for, example, by hydrolysis or
enzymatic cleavage, e.g., by proteases. The delivery system may be,
for example, a naturally occurring or synthetic polymer or
copolymer, for example in the form of a hydrogel. Exemplary
polymers with cleavable linkages include polyesters,
polyorthoesters, polyanhydrides, polysaccharides,
poly(phosphoesters), polyamides, polyurethanes,
poly(imidocarbonates) and poly(phosphazenes).
[0091] The compounds disclosed herein may be administered
enterally, orally, parenterally, sublingually, by inhalation (e.g.
as mists or sprays), rectally, or topically in dosage unit
formulations containing conventional nontoxic pharmaceutically
acceptable carriers, adjuvants, and vehicles as desired. For
example, suitable modes of administration include oral,
subcutaneous, transdermal, transmucosal, iontophoretic,
intravenous, intraarterial, intramuscular, intraperitoneal,
intranasal (e.g. via nasal mucosa), subdural, rectal,
gastrointestinal, and the like, and directly to a specific or
affected organ or tissue. For delivery to the central nervous
system, spinal and epidural administration, or administration to
cerebral ventricles, can be used. Topical administration may also
involve the use of transdermal administration such as transdermal
patches or iontophoresis devices. The term parenteral as used
herein includes subcutaneous injections, intravenous,
intramuscular, intrasternal injection, or infusion techniques. The
compounds are mixed with pharmaceutically acceptable carriers,
adjuvants, and vehicles appropriate for the desired route of
administration. Oral administration is a preferred route of
administration, and formulations suitable for oral administration
are preferred formulations. The compounds described for use herein
can be administered in solid form, in liquid form, in aerosol form,
or in the form of tablets, pills, powder mixtures, capsules,
granules, injectables, creams, solutions, suppositories, enemas,
colonic irrigations, emulsions, dispersions, food premixes, and in
other suitable forms. The compounds can also be administered in
liposome formulations. The compounds can also be administered as
prodrugs, where the prodrug undergoes transformation in the treated
subject to a form which is therapeutically effective. Additional
methods of administration are known in the art.
[0092] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions, may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in propylene glycol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, and
isotonic sodium chloride solution. In addition, sterile, fixed oils
are conventionally employed as a solvent or suspending medium. For
this purpose any bland fixed oil may be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid find use in the preparation of injectables.
[0093] Suppositories for rectal administration of the drug can be
prepared by mixing the drug with a suitable nonirritating excipient
such as cocoa butter and polyethylene glycols that are solid at
room temperature but liquid at the rectal temperature and will
therefore melt in the rectum and release the drug.
[0094] Solid dosage forms for oral administration may include
capsules, tablets, pills, powders, and granules. In such solid
dosage forms, the active compound may be admixed with at least one
inert diluent such as sucrose, lactose, or starch. Such dosage
forms may also comprise additional substances other than inert
diluents, e.g., lubricating agents such as magnesium stearate. In
the case of capsules, tablets, and pills, the dosage forms may also
comprise buffering agents. Tablets and pills can additionally be
prepared with enteric coatings.
[0095] Liquid dosage forms for oral administration may include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups, and elixirs containing inert diluents commonly used in the
art, such as water. Such compositions may also comprise adjuvants,
such as wetting agents, emulsifying and suspending agents,
cyclodextrins, and sweetening, flavoring, and perfuming agents.
[0096] The compounds disclosed herein can also be administered in
the form of liposomes. As is known in the art, liposomes are
generally derived from phospholipids or other lipid substances.
Liposomes are formed by mono- or multilamellar hydrated liquid
crystals that are dispersed in an aqueous medium. Any non-toxic,
physiologically acceptable and metabolizable lipid capable of
forming liposomes can be used. The present compositions in liposome
form can contain, in addition to one or more of the compounds
disclosed herein, stabilizers, preservatives, excipients, and the
like. The preferred lipids are the phospholipids and phosphatidyl
cholines (lecithins), both natural and synthetic. Methods to form
liposomes are known in the art. See, for example, Prescott, Ed.,
Methods in Cell Biology, Volume XIV, Academic Press, New York,
N.W., p. 33 et seq (1976).
[0097] The invention also provides articles of manufacture and kits
containing materials useful for treating or suppressing
mitochondrial diseases. The article of manufacture comprises a
container with a label. Suitable containers include, for example,
bottles, vials, and test tubes. The containers may be formed from a
variety of materials such as glass or plastic. The container holds
a composition having an active agent which is effective for
treating or suppressing mitochondrial diseases. The active agent in
the composition is one or more of the compounds disclosed herein.
The label on the container indicates that the composition is used
for treating or suppressing mitochondrial diseases, and may also
indicate directions for either in vivo or in vitro use, such as
those described above.
[0098] The invention also provides kits comprising any one or more
of the compounds disclosed herein. In some embodiments, the kit of
the invention comprises the container described above. In other
embodiments, the kit of the invention comprises the container
described above and a second container comprising a buffer. It may
further include other materials desirable from a commercial and
user standpoint, including other buffers, diluents, filters,
needles, syringes, and package inserts with instructions for
performing any methods described herein.
[0099] In other aspects, the kits may be used for any of the
methods described herein, including, for example, to treat an
individual with a mitochondrial disorder, or to suppress a
mitochondrial disorder in an individual.
[0100] The amount of active ingredient that may be combined with
the carrier materials to produce a single dosage form will vary
depending upon the host to which the active ingredient is
administered and the particular mode of administration. It will be
understood, however, that the specific dose level for any
particular patient will depend upon a variety of factors including
the activity of the specific compound employed, the age, body
weight, body area, body mass index (BMI), general health, sex,
diet, time of administration, route of administration, rate of
excretion, drug combination, and the type, progression, and
severity of the particular disease undergoing therapy (or of the
energy biomarker being modulated). The pharmaceutical unit dosage
chosen is usually fabricated and administered to provide a defined
final concentration of drug in the blood, tissues, organs, or other
targeted region of the body. The therapeutically effective amount
or effective amount for a given situation can be readily determined
by routine experimentation and is within the skill and judgment of
the ordinary clinician.
[0101] Examples of dosages which can be used are an effective
amount within the dosage range of about 0.1 .mu.g/kg to about 300
mg/kg, or within about 1.0 .mu.g/kg to about 40 mg/kg body weight,
or within about 1.0 .mu.g/kg to about 20 mg/kg body weight, or
within about 1.0 .mu.g/kg to about 10 mg/kg body weight, or within
about 10.0 .mu.g/kg to about 10 mg/kg body weight, or within about
100 .mu.g/kg to about 10 mg/kg body weight, or within about 1.0
mg/kg to about 10 mg/kg body weight, or within about 10 mg/kg to
about 100 mg/kg body weight, or within about 50 mg/kg to about 150
mg/kg body weight, or within about 100 mg/kg to about 200 mg/kg
body weight, or within about 150 mg/kg to about 250 mg/kg body
weight, or within about 200 mg/kg to about 300 mg/kg body weight,
or within about 250 mg/kg to about 300 mg/kg body weight. Other
dosages which can be used are about 0.01 mg/kg body weight, about
0.1 mg/kg body weight, about 1 mg/kg body weight, about 10 mg/kg
body weight, about 20 mg/kg body weight, about 30 mg/kg body
weight, about 40 mg/kg body weight, about 50 mg/kg body weight,
about 75 mg/kg body weight, about 100 mg/kg body weight, about 125
mg/kg body weight, about 150 mg/kg body weight, about 175 mg/kg
body weight, about 200 mg/kg body weight, about 225 mg/kg body
weight, about 250 mg/kg body weight, about 275 mg/kg body weight,
or about 300 mg/kg body weight. Compounds disclosed herein may be
administered in a single daily dose, or the total daily dosage may
be administered in divided dosage of two, three or four times
daily.
[0102] While the compounds disclosed herein can be administered as
the sole active pharmaceutical agent, they can also be used in
combination with one or more other agents used in the treatment or
suppression of disorders. Representative agents useful in
combination with the compounds disclosed herein for the treatment
or suppression of mitochondrial diseases include, but are not
limited to, Coenzyme Q, vitamin E, idebenone, MitoQ, vitamins, and
antioxidant compounds.
[0103] When additional active agents are used in combination with
the compounds disclosed herein, the additional active agents may
generally be employed in therapeutic amounts as indicated in the
Physicians' Desk Reference (PDR) 53rd Edition (1999), which is
incorporated herein by reference, or such therapeutically useful
amounts as would be known to one of ordinary skill in the art.
[0104] The compounds disclosed herein and any other therapeutically
active agents can be administered at the recommended maximum
clinical dosage or at lower doses. Dosage levels of the active
compounds in the compositions disclosed herein may be varied so as
to obtain a desired therapeutic response depending on the route of
administration, severity of the disease and the response of the
patient. When administered in combination with other therapeutic
agents, the therapeutic agents can be formulated as separate
compositions that are given at the same time or different times, or
the therapeutic agents can be given as a single composition.
[0105] The invention is further illustrated by means of the
following examples, which are not intended to limit the invention
in any manner.
EXAMPLES
Example 1
[0106] ##STR10##
[0107] Step 1: A 2 L 3-N flask was charged with
2,3,5-trimethyl-benzene-1,4-diol (201; 50 g, 0.33 mol) and MEK (750
mL) to yield an amber solution. Potassium carbonate (210 g, 1.64
mol) was charged to the solution. After 30 min at room temperature,
MeI (81.2 mL, 1.31 mol) was added to the brown suspension. The
reaction mixture was heated to 65.degree. C. for 72 h. After
cooling to room temperature, the reaction mixture was concentrated
to dryness by rotary evaporation to give a white paste. The paste
was washed with EtOAc (3.times.300 mL). The EtOAc extracts were
combined and concentrated by rotary evaporation. The resulting
yellow-brown oil was chromatographed (80:20/heptanes:EtOAc) to
yield 1,4-dimethoxy-2,3,5-trimethyl-benzene (47.2 g, 80%). .sup.1H
NMR (400 MHz; CDCl.sub.3; ppm): 6.55 (s, 1H), 3.80 (s, 3H), 3.68
(s, 3H), 2.30 (s, 3H), 2.22 (s, 3H), 2.14 (s, 3H).
[0108] Step 2: A flask was charged with
1,4-dimethoxy-2,3,5-trimethyl-benzene (47.2 g, 0.26 mol), glacial
acetic acid (250 mL), and paraformaldehyde (39.3 g, 1.31 mol) to
yield a yellow suspension. Anhydrous HCl gas was then slowly
bubbled through the reaction mixture for 1.5 h producing a clear
amber solution. The reaction mixture was then diluted with water
(300 mL) and extracted with MTBE (3.times.300 mL). The combined
MTBE layers were dried over Na.sub.2SO.sub.4, filtered and
concentrated by rotary evaporation. Purification of the crude
product by silica gel chromatography (95:5/heptanes:EtOAc) yielded
1-chloromethyl-2,5-dimethoxy-3,4,6-trimethyl-benzene (202; 48.7 g,
81%). .sup.1H NMR (400 MHz; CDCl.sub.3; ppm): 4.76 (s, 2H), 3.81
(s, 3H), 3.68 (s, 3H), 2.36 (s, 3H), 2.23 (s, 3H), 2.21 (s,
3H).
[0109] Typical Procedure For Kochi Coupling: A 100 mL 3-N flask (A)
was inerted and charged with
1-chloromethyl-2,5-dimethoxy-3,4,6-trimethyl-benzene (202; 3 g,
13.1 mmol) and degassed THF (30 mL). The flask was then cooled to
0.degree. C. A separate 100 mL 3-N flask (B) was inerted and
charged with the appropriate alkyl Grignard reagent (17.1 mmol).
Flask B was then cooled to 0.degree. C. A third 50 mL flask (C) was
inerted and charged with copper (II) chloride (88 mg, 0.66 mmol),
lithium chloride (56 mg, 1.32 mmol) and degassed THF (15 mL). After
5 min., the rusty orange solution in flask C was transferred to the
solution of 1-chloromethyl-2,5-dimethoxy-3,4,6-trimethyl-benzene in
flask A. The contents of flask A were then transferred dropwise via
syringe to the Grignard solution in flask B over 30 min
(exothermic). The reaction was allowed to stir for 16 h. The
reaction was quenched with MTBE (20 mL) and saturated aqueous
NH.sub.4Cl (20 mL). After stirring for 10 min., the resulting
suspension was filtered to remove dimerized side product. The
aqueous layer was extracted with MTBE (3.times.20 mL). The combined
MTBE layers were concentrated by rotary evaporation to yield a
white residue. The residue was purified by silica gel
chromatography (1:1/DCM:heptane) to yield desired coupled products
(see 203, 204). ##STR11##
[0110] Using an n-pentyl Grignard reagent,
1-hexyl-2,5-dimethoxy-3,4,6-trimethyl-benzene (203) was synthesized
(38%, clear colorless oil); .sup.1H NMR (400 MHz; CDCl.sub.3; ppm):
3.69 (s, 1H), 3.66 (s, 1H), 2.63-2.59 (m, 2H), 2.24 (s, 3H), 2.20
(s, 6H), 1.53-1.28 (m, 8H), 0.90 (t, J=7.1 Hz, 3H). ##STR12##
[0111] Using an n-heptyl Grignard reagent,
1-octyl-2,5-dimethoxy-3,4,6-trimethyl-benzene (204) was synthesized
(57%, clear colorless oil); .sup.1H NMR (400 MHz; CDCl.sub.3; ppm):
3.71 (s, 1H), 3.68 (s, 1H), 2.65-2.61 (m, 2H), 2.25 (s, 3H), 2.21
(s, 6H), 1.53-1.31 (m, 12H), 0.92 (t, J=7.1 Hz, 3H). ##STR13##
[0112] CAN Oxidation: A flask was charged with
1-hexyl-2,5-dimethoxy-3,4,6-trimethyl-benzene (203; 1.75 g, 7.5
mmol) and CAN (20 mL) then cooled to 0.degree. C. A solution of CAN
(8.4 g, 15.4 mmol) in water (10 mL) was added to the flask. After 1
h the reaction was complete. The reaction mixture was extracted
with MTBE (3.times.20 mL). The combined MTBE layers were dried over
MgSO.sub.4, filtered and concentrated by rotary evaporation to
yield 2-hexyl-3,5,6-trimethyl-[1,4]benzoquinone (205) as a
yellow-orange oil which solidified upon standing (1.64 g, 88%).
.sup.1H NMR (400 MHz; CDCl.sub.3; ppm) 2.49-2.46 (m, 2H), 2.04 (s,
3H), 2.03 (s, 6H), 1.44-1.22 (m, 8H), 0.90 (t, J=7.1 Hz, 3H).
##STR14##
[0113] A flask was charged with
1-octyl-2,5-dimethoxy-3,4,6-trimethyl-benzene (204; 1.75 g, 7.5
mmol) and CAN (20 mL) then cooled to 0.degree. C. A solution of CAN
(8.4 g, 15.4 mmol) in water (10 mL) was added to the flask. After 1
h the reaction was complete. The reaction was diluted with water
(50 mL) and the yellow precipitate was filtered and washed with
water (20 mL). The fine yellow needles were dried under high vacuum
to give pure 2-octyl-3,5,6-trimethyl-[1,4]benzoquinone (206; 1.69
g, 86%). .sup.1H NMR (400 MHz; CDCl.sub.3; ppm): 2.49-2.45 (m, 2H),
2.04 (s, 3H), 2.03 (s, 6H), 1.45-1.19 (m, 12H), 0.89 (t, J=6.2 Hz,
3H).
Example 2
Decarboxylative Coupling
Example 2A
[0114] ##STR15##
[0115] To a 250 ml round bottom flask was added
2,3,5-trimethyl-[1,4]benzoquinone (1.50 g, 9.98 mmole), linolenic
acid (2.94 g, 10.4 mmole), and silver nitrate (1.83 g, 10.8 mmole)
in a 1:1 mixture of water and acetonitrile (100 ml). The solution
was heated to 70.degree. C. under argon and an aqueous solution of
K.sub.2S.sub.2O.sub.8 (2.55 g, 11.5 mmole in 50 ml water) was added
dropwise to the homogenous solution over 2.5 hours using a syringe
pump. The reaction mixture was allowed to stir an additional 30
minutes at 70.degree. C., then cooled to room temperature. To the
mixture was added MTBE (200 ml) and water (100 ml). The organic
layer was separated and washed with saturated NaHCO.sub.3 (100 ml),
then brine (2.times.200 ml). The MTBE solution was dried over
sodium sulfate then concentrated to a yellow oil. The crude
product, which contained residual unreacted starting quinone by
TLC, was further purified by silica gel chromatography (120 g,
0-30% EtOAc:heptane) to give pure
2-heptadeca-8,11-dienyl-3,5,6-trimethyl-[1,4]benzoquinone (210;
0.474 g, 12.3%) as a yellow oil. .sup.1H NMR (400 MHz;
d.sub.6-DMSO; ppm): 5.36-5.26 (m, 4H), 2.73 (t, J=5.6 Hz, 2H),
2.42-2.38 (m, 2H), 2.02-1.93 (m, 4H). 1.95 (s, 3H), 1.93 (s, 6H).
1.34-1.22 (m, 16H), 0.84 (t, J=7.2 Hz, 3H).
Example 2B
[0116] ##STR16##
[0117] To a 250 ml round bottom flask was added
2,3,5-trimethyl-[1,4]benzoquinone (0.51 g, 3.4 mmole), oleic acid
(1.0 g, 3.5 mmole), and silver nitrate (0.62 g, 3.6 mmole) in a 1:1
mixture of water and acetonitrile (100 ml). The solution was heated
to 70.degree. C. under argon and an aqueous solution of
K.sub.2S.sub.2O.sub.8 (0.86 g, 3.9 mmole in 50 ml water) was added
dropwise to the homogenous mixture over 2.5 hours using a syringe
pump. The reaction was stirred at 70.degree. C. for an additional
30 minutes after addition, then cooled to room temperature. To the
reaction mixture was added MTBE (200 ml) and water (100 ml). The
organic layer was separated, washed with water (2.times.100 ml),
then brine (2.times.100 ml). The solution was dried over sodium
sulfate and concentrated to a yellow oil. The crude product was
further purified by silica gel chromatography (120 g, 0-30%
EtOAc:heptane) to give pure
2-heptadec-8-enyl-3,5,6-trimethyl-[1,4]benzoquinone (211; 25.2 mg,
2%) as a yellow oil. .sup.1H NMR (400 MHz; d.sub.6-DMSO; ppm):
5.32-5.30 (m 2H), 2.42-2.38 (m, 2H), 1.98-1.93 (m, 4H), 1.95 (s,
3H), 1.93 (s, 6H), 1.40-1.22 (m, 22H), 0.84 (t, J=7.2 Hz, 3H).
Example 2C
[0118] ##STR17##
[0119] Step 1: To a 500 ml round bottom flask equipped with a
stir-bar was added 2-tert-butyl-6-methyl-benzene-1,4-diol (212; 18
g, 100 mmole), paraformaldehyde (3.0 g, 100 mmole), SnCl.sub.2
(47.4 g, 250 mmole), DME (200 ml), and concentrated HCl (50 ml,
35%). The flask was fitted with a reflux condenser and the reaction
mixture was heated to 75.degree. C. After 24 hours, the mixture was
cooled to room temperature. To the mixture was added MTBE (300 ml).
The organic fraction was separated and washed with water
(3.times.500 ml) followed by brine (2.times.200 ml). The organic
fraction was dried over sodium sulfate and concentrated to a
red-brown foam. The resultant crude
5-tert-butyl-2,3-dimethyl-benzene-1,4-diol was taken directly to
the next step with no further purification.
[0120] Step 2: To a 500 ml round bottom flask equipped with a
stir-bar was added crude 5-tert-butyl-2,3-dimethyl-benzene-1,4-diol
as a solution in MeCN (200 ml). To the stirring solution at room
temperature was added CAN (114 g, 220 mmole) as a solution in water
(200 ml) in one portion. The biphasic reaction mixture was stirred
vigorously at room temperature for one hour, after which time no
further reaction was detected by TLC analysis (20% EtOAc:heptane).
The reaction mixture was poured into MTBE (500 ml). The organic
layer was separated, then washed with water until the aqueous phase
remained colorless (3.times.200 ml). The solution was then washed
with brine (2.times.200 ml), dried over sodium sulfate, and
concentrated to a red oil. A portion of the crude product was
further purified by silica gel chromatography (120 g, 0-20%
EtOAc:heptane) to give 5-tert-butyl-2,3-dimethyl-[1,4]benzoquinone
(213) as a volatile yellow oil. .sup.1H NMR (400 MHz;
C.sub.6D.sub.6; ppm): 6.42 (s, 1H), 1.66 (q, J=1.2 Hz, 3H), 1.61
(q, J=1.2 Hz, 3H), 1.12 (s, 9H).
[0121] Step 3: To a 250 ml round bottom flask equipped with a
stir-bar was added 5-tert-butyl-2,3-dimethyl-[1,4]benzoquinone
(213; 0.68 g, 3.5 mmole), heptanoic acid (0.49 g, 3.7 mmole),
AgNO.sub.3 (0.64 g, 3.8 mmole), acetonitrile (50 ml) and water (50
ml). The fully homogenous solution was heated to 70.degree. C.
under argon while an aqueous solution of K.sub.2S.sub.2O.sub.8
(0.91 g, 4.1 mmole in 30 ml water) was added dropwise over 2.5
hours using a syringe pump. The reaction mixture was allowed to
stir an additional 30 minutes at 70.degree. C., then cooled to room
temperature. To the mixture was added heptane (100 ml) and water
(100 ml). The organic layer was separated and washed with saturated
NaHCO.sub.3 (1.times.50 ml) followed by brine (2.times.100 ml). The
organic layer was dried over sodium sulfate and concentrated to a
yellow oil. The crude product was further purified by preparative
TLC (silica gel: 200.times.200.times.2 mm; 100% heptane loading; 5%
EtOAc:heptane elution). The fastest running bands, as visualized by
UV, were excised, extracted from the silica gel with methyl
tert-butyl ether, and the extract concentrated to a yellow oil to
give a yellow oil. The residue was further purified by flash
chromatography [silica gel: 40 g; 0-20% 100% heptane loading; 0-20%
EtOAc/heptane gradient elution] to give pure
2-tert-butyl-3-hexyl-5,6-dimethyl-[1,4]benzoquinone (214; 83 mg,
8.4%) as a yellow oil. .sup.1H NMR (400 MHz; C.sub.6D.sub.6; ppm):
2.71-2.67 (m, 2H), 1.67-1.66 (m, 6H), 1.51-1.24 (m, 8H), 1.37 (s,
9H), 0.88 (t, J=7.2 Hz, 3H).
Example 2D
[0122] ##STR18##
[0123] Step 1: To a 500 ml round bottom flask equipped with a
stir-bar was added 2,6-diisopropyl-benzene-1,4-diol (215; 5.0 g, 26
mmole), paraformaldehyde (0.78 g, 26 mmole), SnCl.sub.2 (18.9 g,
100 mmole), diisopropyl ether (200 ml), and concentrated HCl (60
ml, 35%). The flask was fitted with a reflux condenser and the
reaction mixture was heated to 66.degree. C. After 24 hours, the
mixture was cooled to room temperature (the reaction remained
biphasic throughout. To the reaction mixture was added MTBE (200
ml). The organic fraction was separated and washed with HCl
solution (1.times.200 ml, 1N), water (3.times.100 ml), and brine
(2.times.100 ml). The organic fraction was dried over sodium
sulfate and concentrated to yellow oil. The resultant crude
2,6-diisopropyl-3-dimethyl-benzene-1,4-diol was taken directly to
the next step with no further purification.
[0124] Step 2: To a 500 ml round bottom flask equipped with a
stir-bar was added crude
2,6-diisopropyl-3-dimethyl-benzene-1,4-diol as a solution in MeCN
(100 ml). To the stirring solution at room temperature was added
CAN (28.5 g, 55.0 mmole) as a solution in water (100 ml) in one
portion. The biphasic reaction mixture was stirred vigorously at
room temperature for one hour, after which time no further reaction
was detected by TLC analysis (20% EtOAc:heptane). The reaction
mixture was poured into MTBE (200 ml). The organic layer was
separated, then washed with water (2.times.100 ml). The solution
was then washed with brine (2.times.100 ml), dried over sodium
sulfate, and concentrated to a red-yellow oil. The crude product
was further purified by silica gel chromatography (0-5%
EtOAc:heptane) to give 3,5-diisopropyl-2-methyl-[1,4]benzoquinone
(216) as a volatile yellow oil. .sup.1H NMR (400 MHz;
C.sub.6D.sub.6; ppm): 6.30 (d, J=1.4 Hz, 1H), 2.94-2.91 (m, 1H),
2.85-2.81 (m, 1H), 1.80 (d, J=1.2 Hz, 3H), 1.16 (d, J=6.8 Hz, 6H),
0.81 (dd, J=1.4 Hz, J.sub.2=6.4 Hz, 6H).
[0125] Step 3: To a 250 ml round bottom flask equipped with a
stir-bar was added 3,5-diisopropyl-2-methyl-[1,4]benzoquinone (216;
1.03 g, 5.00 mmol), linoleic acid (1.63 ml, 1.47 g, 5.24 mmol),
silver(I) nitrate (917 mg, 5.40 mmol), acetonitrile (35 ml), and
water (25 ml). The solution was heated under balloon-closed ambient
atmosphere to 75.degree. C., at which it was homogenous. Potassium
persulfate (1.28 g, 5.75 mmol) in water (30 ml) was then added
dropwise over 4 hours via syringe pump. Following complete addition
the reaction mixture was heated for a further 2 hours and then the
reaction volume reduced by approximately half under reduce pressure
on a rotary evaporator. Water (50 ml) was added to the concentrate
and the mixture extracted with MTBE (3.times.50 ml). The combined
organics were washed with brine (50 ml), dried (sodium sulfate),
and concentrated to a yellow oil. A portion of the crude product
was further purified by preparative TLC (silica gel:
200.times.200.times.2 mm; 100% heptane loading; 5% MTBE:heptane
elution). The fastest running bands, as visualized by UV, were
excised, extracted from the silica gel with MTBE, and the extract
concentrated to a yellow oil to give a yellow oil (280 mg). The
residue was further purified by flash chromatography [silica gel:
40 g; 0-20% 100% heptane loading; 0-20% EtOAc/heptane gradient
elution] to give
2-heptadeca-8,11-dienyl-3,5-diisopropyl-6-methyl-[1,4]benzoquinone
(217; 65.6 mg, 2.9% mass yield) as a yellow oil which was pure as
determined by reverse-phase HPLC. .sup.1H NMR (400 MHz;
d.sub.6-DMSO; ppm): 5.40 (m, 4H), 3.05-2.94 (m, 2H), 2.73 (t, J=5.6
Hz, 2H), 2.47-2.30 (m, 2H), 2.02-1.96 (m, 4H), 1.95 (s, 3H),
1.29-1.20 (m, 16H), 1.21 (d, J=6.8 Hz, 6H), 1.19 (d, J=7.2 Hz, 6H),
0.84 (t, J=7.2 Hz, 3H).
Example 2E
[0126] ##STR19##
[0127] To a 100 ml round bottom flask equipped with a stir-bar was
added 3,5-diisopropyl-2-methyl-[1,4]benzoquinone (216, see Example
2D; 1.03 g, 5.00 mmol), octanoic acid (83211, 757 mg, 5.24 mmole),
silver (I) nitrate (917 mg, 5.40 mmol), acetonitrile (35 ml), and
water (25 ml). The solution was heated under balloon-closed ambient
atmosphere to 75.degree. C. and was homogenous. Potassium
persulfate (1.28 g, 5.75 mmol) in water (30 ml) was then added
dropwise over 4 hours via syringe pump. Following complete addition
the reaction mixture was heated for a further 2 hours and then the
reaction volume reduced by approximately half under reduce pressure
on a rotary evaporator. Water (50 ml) was added to the concentrate
and the mixture extracted with MTBE (3.times.50 ml). The combined
organics were washed with brine (50 ml), dried (sodium sulfate),
and concentrated to a yellow oil (1.2 g). Approx. 75% of the
residue was purified in 150-200 mg portions by preparative-TLC
[silica gel: 200.times.200.times.2 mm; 100% heptane loading; 5%
ethyl acetate/heptane elution]. The fastest running bands, as
visualized by UV, were combined, extracted from the silica gel with
MTBE, and the extract concentrated to a yellow oil (.about.300 mg).
The residue was further purified by flash chromatography [silica
gel: 120 g; 100% heptane loading; 3-6% ethyl acetate/heptane
gradient elution] to give the
2-heptyl-3,5-diisopropyl-6-methyl-[1,4]benzoquinone (218) as a
bright yellow oil (288 mg, 21% mass yield). .sup.1H NMR (400 MHz;
C.sub.6D.sub.6; ppm): 2.93-2.80 (m, 2H), 2.49-2.46 (m, 2H), 1.84
(s, 3H), 1.43-1.18 (m, 10H), 1.33 (d, J=6.8 Hz, 6H), 1.19 (d, J=6.8
Hz, 6H), 0.88 (t, J=7.2 Hz, 3H).
[0128] The disclosures of all publications, patents, patent
applications and published patent applications referred to herein
by an identifying citation are hereby incorporated herein by
reference in their entirety.
[0129] Although the foregoing invention has been described in some
detail by way of illustration and example for purposes of clarity
of understanding, it is apparent to those skilled in the art that
certain minor changes and modifications will be practiced.
Therefore, the description and examples should not be construed as
limiting the scope of the invention.
* * * * *